Ivyiyumviro bitandukanye vy’ukwikingira ibimera biratanga ubuyobozi buhambaye bwo gusigura ingene ibimera bihindura ibiribwa, ariko ivyo bivuga bihambaye biracariho kugira ngo bigeragezwe. Aha, twakoresheje isesengura ry’uburemere bw’ibintu (MS/MS) kugira ngo dusuzume mu buryo bubereye ivy’imiterere y’ibimera vy’itabi biva ku bimera bimwebimwe gushika ku bantu n’ubwoko bufitaniye isano rya hafi, kandi dukora umubare munini w’ivyiyumviro vy’ibiranga uburemere bw’ibintu bishingiye ku bimenyetso vy’ibintu bihurikiye hamwe biri mu makuru. Inzego zo kugerageza ivyiyumviro nyamukuru vy’ukwikingira neza (OD) n’ivyiyumviro vy’intumbero yimuka (MT). Igice c’amakuru c’imiterere y’ibimera gihuye n’inyigisho ya OD, ariko kivuguruza ubuhanuzi nyamukuru bw’inyigisho ya MT ku bijanye n’imigenderanire y’ibimera iterwa n’ibikoko birya ivyatsi. Kuva ku rugero rw’iterambere ry’ibikoko bitobito gushika ku rugero runini, ikimenyetso ca jasmonate caramenyekanye ko ari co kintu nyamukuru kigaragaza OD, mu gihe ikimenyetso ca ethylene catanga ugutunganya neza inyishu y’ibikoko birya ivyatsi ivugwa n’uruzitiro rw’ibihimba vy’umubiri rwa MS/MS.
Ivyo bihimba bidasanzwe bihindura ibiribwa bifise imiterere itandukanye ni vyo bifasha cane mu gutuma ibimera bimenyerera ibidukikije, cane cane mu kwikingira abansi (1). Ubutandukane butangaje bw'imihindukire idasanzwe iboneka mu bimera bwatumye habaho ubushakashatsi bwimbitse mu myaka mirongo ku mice myinshi y'ibikorwa vyavyo mu bidukikije, kandi bwatumye haba urutonde rurerure rw'ivyiyumviro vy'ukwikingira ibimera, arivyo iterambere ry'ibidukikije n'iterambere ry'imigenderanire hagati y'ibimera n'udukoko. Ubushakashatsi bushingiye ku bintu vy’ukuri buratanga ubuyobozi buhambaye (2). Ariko rero, izo nyigisho zo kwikingira ibimera ntizakurikiye inzira isanzwe y’ugutekereza ku vyo umuntu abona, aho ivyiyumviro nyamukuru vyari ku rugero rumwe rw’isesengura (3) kandi vyageragejwe mu buryo bw’igerageza kugira ngo bitere imbere mu nzira ikurikira y’iterambere ry’ivyiyumviro (4). Ivyiyumviro vy’ubuhinga bibuza gukusanya amakuru ku mice yihariye y’ibinyabuzima kandi ntibibemwo isesengura ry’ibinyabuzima vyihariye, gutyo bikabuza ukugereranya hagati y’ibice bihambaye mu guteza imbere ivyiyumviro (5). Kubura amakuru yuzuye y’ivy’imibiri n’amahera asanzwe yo kugereranya ibikorwa vyo gutunganya ikibanza c’imibiri hagati y’imigwi itandukanye y’ibimera birabuza gukura kw’ubuhinga bw’ivyo bikorwa.
Ivyiza bishasha vyakozwe mu bijanye n’ubuhinga bwo gupima uburemere bw’ibinyabuzima (MS/MS) birashobora gusobanura neza ihinduka ry’ibinyabuzima hagati y’ubwoko bw’ibinyabuzima bimwe bimwe, kandi bishobora gufatanywa n’uburyo bwo guharura kugira ngo umuntu abare ukuntu ivyo bivange bikomeye bisa. Ubumenyi bubanza bw’ivy’ubuhinga bwa shimi (5). Ihuriro ry’ubuhinga buteye imbere mu gusesangura no mu gukoresha ubuhinga bwa none ritanga urutonde rukenewe rwo kugerageza igihe kirekire ibintu vyinshi vyavuzwe n’inyigisho z’ibidukikije n’ivy’iterambere ry’ubudasa bw’ibinyabuzima. Shannon (6) yashizeho inyigisho y’amakuru ku ncuro ya mbere mu ngingo yiwe y’intango mu 1948, ashiraho umushinge w’ugusesangura amakuru mu mibare, iyo nyigisho ikaba yarakoreshejwe mu bindi bikorwa vyinshi uretse uko yakoreshwa mu ntango. Mu vy’ubuhinga bw’ivy’imitsi, inyigisho y’amakuru yarakoreshejwe neza mu gupima amakuru y’urutonde (7). Mu bushakashatsi bw’ivy’inyandiko, inyigisho y’amakuru isesangura amahinduka yose aba mu nyandiko (8). Mu bushakashatsi bwa kera, twakoresheje urutonde rw’imibare rw’inyigisho y’amakuru ku vy’ubuhinga bw’imibiri kugira ngo tudondore ubuhinga bw’imibiri y’urugero rw’imitsi mu bimera (9). Aha, dufatanya urugendo rw’ibikorwa rushingiye kuri MS/MS n’inzego z’imibare z’inyigisho y’amakuru, irangwa n’ubudasa bw’ibifungurwa mu mafaranga rusangi, kugira ngo tugereranye ivyiyumviro nyamukuru vy’inyigisho y’ukwikingira ibimera y’ibifungurwa vy’ibinyabuzima biterwa n’ibikoko birya ivyatsi.
Ivyiyumviro vy’ukwikingira ibimera akenshi birahuza kandi bishobora guca mu mice ibiri: ivyo bigerageza gusigura ukuntu ibimera bihinduka bivanye n’ibikorwa vy’ukwikingira, nk’ukwikingira neza (OD) (10), intumbero yigendagenda (MT) (11) ) N’ukuntu (12) uburyo buhinduka (12) theory of mechanisments in, ukuboneka biragira ico bikoze ku gukura kw’ibimera no ku kwirundanira kw’ibintu bidasanzwe bihindura ibiribwa, nk’ivyo bita carbone: ivyiyumviro vy’uburinganire bw’ingaburo (13), ivyiyumviro vy’urugero rw’ukukura (14 ), n’ivyiyumviro vy’uburinganire bw’ukukura n’ugutandukanya (15). Ivyo vyiyumviro bibiri biri ku rwego rutandukanye rw’isesengura (4). Ariko rero, inyigisho zibiri zijanye n’ibikorwa vyo kwikingira ku rugero rw’ibikorwa ni zo ziganza ikiyago ku bijanye n’ukwikingira kw’ibimera n’ukwikingira kw’ibimera: inyigisho ya OD, yiyumvira ko ibimera bishiramwo amahera mu kwikingira kwavyo guzimvye gusa igihe bikenewe, nk’akarorero, igihe binywanywa Igihe igikoko c’ivyatsi gitera, rero, hakurikijwe ubushobozi bwo kwikingira igikorwa gihabwa (10); MT hypothesis ivuga ko ata nzira y’ihinduka ry’ibinyabuzima bihinduka, ariko ibinyabuzima bihinduka mu buryo butari bwo, gutyo bikaba bishoboka Guhagarika “intumbero y’inyigisho” y’ibinyabuzima vyo gutera ibikoko birya ivyatsi. Mu yandi majambo, izo nyigisho zibiri zitanga ubuhanuzi bunyuranye ku bijanye n’uguhinduka kw’imibiri bishika inyuma y’igitero c’ibikoko birya ivyatsi: isano hagati y’ukwirundanira kw’imibiri mu buryo bumwe bifise igikorwa co kwikingira (OD) n’ihinduka ry’imibiri ritagira inzira (MT) (11 ).
Ivyiyumviro vya OD na MT ntibirimwo gusa amahinduka aterwa n’ibinyabuzima, ariko kandi n’ingaruka z’ibidukikije n’iterambere z’ukwirundanira kw’ivyo binyabuzima, nk’ibiciro vyo kwimenyereza n’ivyiza vy’ayo mahinduka y’ibinyabuzima mu bidukikije vyihariye (16). Naho ivyo vyiyumviro vyose vyemeza igikorwa co kwikingira c’ibinyabuzima bidasanzwe, bishobora kuba bizimvye canke bitazimvye, ubuhanuzi nyamukuru butandukanya ivyiyumviro vya OD na MT buri mu nzira y’amahinduka y’ibinyabuzima aterwa n’ibinyabuzima. Ubuhanuzi bw’inyigisho ya OD ni bwo bwaronse ivyiyumviro vy’igerageza kuruta ibindi vyose gushika ubu. Ivyo bipimo birimwo inyigisho y’ibikorwa vy’ukwikingira bitaziguye canke bitaziguye vy’imitsi itandukanye y’ibintu vyihariye biri mu nzu z’ivyatsi n’ibidukikije, hamwe n’ihinduka ry’inzira y’iterambere ry’ibimera (17-19). Ariko rero, gushika ubu, kubera ukubura urutonde rw’ibikorwa n’imibare y’isesengura ry’isi yose ry’ubudasa bw’imibiri y’ikinyabuzima ico ari co cose, ubuhanuzi nyamukuru bw’itandukaniro hagati y’izo nyigisho zibiri (ni ukuvuga uburongozi bw’imihindukire y’imibiri) buracariho kugira ngo bugeragezwe. Aha, turatanga mwene ukwo gusesangura.
Kimwe mu biranga cane ibinyabuzima bihindura ibiribwa mu bimera ni ubudasa bwavyo burengeje urugero ku rwego rwose kuva ku bimera bimwe bimwe, ku bimera bimwe bimwe gushika ku bwoko busa (20). Impinduka nyinshi z’ingero mu bihimba vy’umubiri bidasanzwe zishobora kubonwa ku rugero rw’abantu, mu gihe itandukaniro rikomeye ry’uburyo rikunda kugumaho ku rugero rw’ubwoko (20). Ku bw’ivyo, ubudasa bw’ibinyabuzima ni co kintu nyamukuru kigaragaza ubudasa bw’ibikorwa, bugaragaza ubushobozi bwo kwimenyereza mu bibanza bitandukanye, cane cane ivyo bibanza bifise ubushobozi butandukanye bwo guterwa n’udukoko tudasanzwe be n’ibikoko birya ivyatsi bisanzwe (21). Kuva mu ngingo ya Fraenkel (22) y’intangamarara yerekeye imvo zituma habaho ibintu bihindura ibiribwa mu bimera, gukorana n’udukoko dutandukanye vyabonwa ko ari ibintu bihambaye bitera ingorane mu guhitamwo, kandi ivyo bikorana vyemerwa ko ari vyo vyahinduye ibimera mu gihe c’ubwihindurize. Inzira y’imibiri (23). Ubutandukane hagati y’ubwoko mu bwoko bw’ibinyabuzima bidasanzwe bushobora kandi kwerekana uburinganire bw’umubiri bujanye n’ukwikingira kw’ibimera ku ngabire z’ibikoko birya ivyatsi, kuko ubwo bwoko bubiri akenshi bufitaniye isano ribi (24). Naho bishobora kuba vyiza kuguma umuntu afise uburinzi bwiza ibihe vyose, amahinduka y’imibiri ajanye n’uburinzi ashika ku gihe aratanga inyungu zigaragara mu kwemera ko ibiterwa bishobora gutanga ubutunzi bw’agaciro ku bindi bikoresho vy’umubiri (19, 24), no kwirinda gukenera gukorana n’ibindi binyabuzima. Ivyonona vy’ingwati (25). Ikindi, ivyo bihinduka vy’ibinyabuzima bidasanzwe biterwa n’udukoko birya ivyatsi bishobora gutuma haba ugukwiragira gusenyura mu bantu (26), kandi bishobora kwerekana ugusoma gutaziguye kw’amahinduka akomeye y’ibinyabuzima mu kimenyetso c’aside ya jasmonic (JA), bishobora kuguma mu bantu. Ibimenyetso vya JA vyinshi n’ivyo hasi ni uguhuza hagati y’ukwikingira ibikoko birya ivyatsi n’uguhiganwa n’ubwoko bumwe bumwe (27). Ikindi, inzira zidasanzwe z’ibinyabuzima zizotakaza vuba kandi zihinduke mu gihe c’ubwihindurize, ivyo bikaba bizotuma habaho ugusangira kw’ibinyabuzima mu bwoko bufitaniye isano rya hafi (28). Ivyo bihimba bishobora gushingwa vuba cane mu kwishura ku guhinduka kw’imiterere y’ibikoko birya ivyatsi (29), ivyo bikaba bisobanura ko uguhinduka kw’imiryango y’ibikoko birya ivyatsi ari co kintu nyamukuru gituma habaho ubudasa bw’ibifungurwa.
Aha, twatorera umuti vyihariye ingorane zikurikira. (I) Ni gute udukoko turya ivyatsi dusubira gutunganya ivy’ibimera? (Ii) Ni ibihe bihimba nyamukuru vy’amakuru y’ubuhinga bwo guhindura ibiribwa bishobora gupimwa kugira ngo hasuzumwe ivyo ivyiyumviro vy’ukwikingira vy’igihe kirekire bivuga? (Iii) Nimba twosubira gutegura umubiri w’ibimera mu buryo budasanzwe ku muntu atera, nimba ari ukwo biri, ni uruhara uruhe umusemburo w’ibimera ugira mu guhindura inyishu yihariye y’umubiri, kandi ni ibiki bihindura umubiri bifasha mu gutuma ubwoko bw’ibinyabuzima bugira ico bukoze ku bijanye n’ukwikingira? (Iv) Kubera ko ubuhanuzi bwavuzwe n’ivyiyumviro vyinshi vy’ukwigwanira bishobora kwaguka ku nzego zose z’imitsi y’ibinyabuzima, twabajije ingene inyishu y’imibiri iterwa n’ibinyabuzima ihuye kuva ku kugereranya imbere gushika ku kugereranya hagati y’ubwoko? Kugira ngo ivyo bishoboke, twarize mu buryo butunganye ivyerekeye imibiri y’amababi y’itabi, kikaba ari igiterwa c’akarorero c’ibidukikije gifise ubushobozi bwo guhindura imibiri yihariye, kandi kikaba gikora neza ku nkoko z’ibikoko bibiri vy’ivyatsi vy’imvukira, Lepidoptera Datura (Ms) ( Bitera ubukazi cane, ahanini birya) Ku bikoko bita Solanaceae vy’ibikoko vyitwa letora litterap na SpodoS “genus”, n’ibimera vy’abashitsi vy’i Solanaceae n’ibindi bimera vy’abashitsi vy’iyindi miryango n’imiryango Ivyokurya vy’ibitegwa. Twarasuzumye urutonde rw’ibintu bihinduka mu mubiri (MS/MS metabolomics spectrum) maze dukuramwo ibisobanuro vy’imibare vy’inyigisho z’amakuru kugira ngo tugereranye ivyiyumviro vya OD na MT. Gukora amakarata yihariye kugira ngo ugaragaze akaranga k’ibintu nyamukuru bihindura ibiribwa. Iryo sesangura ryaraguwe ku bantu b’imvukira ba N. nasi n’ubwoko bw’itabi bufitaniye isano rya hafi kugira ngo hasuzumwe cane ubudasa buri hagati y’ibimenyetso vy’imisemburo y’ibimera n’ugutera OD.
Kugira ngo dufate ikarita rusangi yerekeye uburemere n’imiterere y’amababi y’itabi ry’ivyatsi, twakoresheje ubuhinga bwo gusesangura no kubara bwari bwateguwe mbere kugira ngo dufate hamwe no gukuraho amakuru y’agaciro kanini yigenga y’imirongo ya MS/MS ivuye mu bihimba vy’ibimera ( 9). Ubwo buryo butagira itandukaniro (bwitwa MS/MS) burashobora kwubaka imirongo y’ibinyabuzima idasubira inyuma, ishobora rero gukoreshwa mu gusesangura kwose kw’urugero rw’ibinyabuzima bivugwa ngaha. Ivyo bihimba vy’ibimera biva mu bihimba vy’umubiri bifise ubwoko butandukanye, birimwo amajana n’ibihumbi vy’ibihimba vy’umubiri (nk’ibice 500-1000-s/MS/MS hano). Aha, turimbura ubuhinga bwo guhindura ibiribwa mu rwego rw’inyigisho y’amakuru, kandi tugapima ubudasa n’ubuhinga bw’ibiribwa bishingiye ku ntunganyo ya Shannon y’ugusangira incuro z’ibiribwa. Dukoresheje uburyo bwakoreshejwe mbere (8), twaharuriye urutonde rw’ibimenyetso bishobora gukoreshwa mu gupima ubudasa bw’ibinyabuzima (ikimenyetso ca Hj), ubuhinga bw’ibinyabuzima (ikimenyetso ca δj) n’ubudasa bw’ibinyabuzima vy’ibinyabuzima bimwe (ikimenyetso ca Si) . Ikindi, twakoresheje urutonde rw’uburemere bw’ibintu (RDPI) kugira ngo tugereranye ubushobozi bw’ibikoko birya ivyatsi (Ishusho 1A) (30). Muri iyo nzira y’imibare, dufata umurongo wa MS/MS nk’igice c’amakuru y’ishimikiro, maze tugakora ubwinshi bw’imirongo ya MS/MS mu ikarita y’ugukwiragira kw’incuro, hanyuma tugakoresha Shannon entropy kugira ngo tugereranye ubudasa bw’ibinyabuzima buva kuri yo. Ubuhinga bwa metabolome bupimwa n’uburyo bwihariye bw’umurongo umwe wa MS/MS. Ku bw’ivyo, ukwiyongera kw’ubwinshi bw’imigwi imwe imwe ya MS/MS inyuma y’uguterwa kw’ibikoko birya ivyatsi bihinduka spectral inducibility, RDPI n’ubuhinga bwihariye, ni ukuvuga ukwiyongera kw’index ya δj, kubera ko habaho ibintu vyinshi vy’umwihariko bihindura ibiribwa kandi hakaba hariho index nini ya Si. Kugabanya urutonde rw’ubudasa bwa Hj vyerekana ko umubare w’ibinyabuzima MS/MS vyavutse ugabanuka, canke ko ugusangira kw’incuro z’imiterere bihinduka mu buryo budahuye cane, mu gihe bigabanya ukudakeka kwayo muri rusangi. Biciye mu kubara urutonde rwa Si, birashoboka ko umuntu agaragaza MS/MS ziterwa n’ibikoko bimwebimwe birya ivyatsi, ahubwo ni MS/MS idasubiza ivy’uguterwa, ari co kimenyetso nyamukuru co gutandukanya ubuhanuzi bwa MT n’ubuhanuzi bwa OD.
(A) Ivyiyumviro vy’imibare bikoreshwa ku bikoko birya ivyatsi (H1 gushika kuri Hx) MS/MS amakuru-inducibility (RDPI), ubudasa (Hj index), ubuhinga (δj index) n’ubuhinga bw’ibinyabuzima (Si index). Ukwongerekana kw’urugero rw’ubuhinga (δj) vyerekana ko, mu mwanya, hazoba hariho ibintu vyinshi vy’ibimera birya ivyatsi, mu gihe ukugabanya ubwoko (Hj) vyerekana ko ibifungurwa bigabanutse canke ko ibifungurwa bigabanuka ku ikarita y’ugusangira . Agaciro ka Si gasuzuma nimba iyo metabolite ari iyo kwihariye ku kintu kinaka (aha, ivyatsi birya) canke bihushanye n’ivyo iguma ku rugero rumwe. (B) Igishushanyo c’ivyiyumviro c’ubuhanuzi bw’inyigisho y’ukwikingira hakoreshejwe umurongo w’inyigisho y’amakuru. Ivyiyumviro vya OD bivuga ko igitero c’ibikoko birya ivyatsi kizokwongerera ubushobozi bwo kwikingira, gutyo bikongera δj. Muri ico gihe nyene, Hj iragabanuka kubera ko urutonde rusubira gutunganirizwa kugira ngo amakuru y’ivy’imibiri agabanuke. Ivyiyumviro vya MT bivuga ko igitero c’ibikoko birya ivyatsi kizotuma habaho amahinduka atagira aho ageze mu bijanye n’imibiri, gutyo bigatuma Hj yongerekana nk’ikimenyetso c’uko amakuru y’imibiri yongerekana kandi bigatuma Si igabanywa mu buryo butari bwo. Twarateye imbere kandi uburyo buvanze, MT nziza cane, aho ibintu bimwebimwe bihindura ibiribwa bifise agaciro kanini ko kwikingira bizokwongereza cane cane (agaciro kanini ka Si), mu gihe ibindi vyerekana inyishu zidasanzwe (agaciro ka Si gatoyi).
Dukoresheje ibisobanuro vy’inyigisho y’amakuru, dusobanura inyigisho ya OD kugira ngo tuvuge imbere y’igihe ko amahinduka yihariye y’ibinyabuzima biterwa n’ibikoko birya ivyatsi mu gihe c’ubuzima butatewe azotuma (i) habaho ukwiyongera kw’ubudasa bw’ibinyabuzima (Si index) bituma ubudasa bw’ibinyabuzima (δj index) Ukwiyongera kw’imigwi imwimwe idasanzwe y’ibinyabuzima bifise agaciro kanini k’ibinyabuzima ( de defense) index) kubera uguhinduka kw’ugukwiragira kw’incuro z’ibifungurwa mu mubiri gushika ku gukwiragira kw’umubiri kwa leptin nyinshi. Ku rugero rw’igihimba kimwe c’umubiri, hariho ugusangira kwa Si ku rutonde, aho igihimba c’umubiri kizokwongerera agaciro ka Si bivanye n’agaciro kaco k’ukwikingira (Ishusho 1B). Kuri uwo murongo, turasigura inyigisho ya MT kugira ngo tubone imbere y’igihe ko ugukangura bizotuma (i) haba amahinduka atagira inzira mu binyabuzima bihindura ibiribwa biva ku kugabanya urutonde rwa δj, na (ii) ukwiyongera kw’urutonde rwa Hj bitewe n’ukwiyongera kw’ukudakeka kw’ibinyabuzima. Canke ubusa, bushobora gupimwa n’ubuhinga bwa Shannon entropy mu buryo bw’ubudasa rusangi. Naho ku bijanye n’imiterere y’ibinyabuzima, inyigisho ya MT izovuga imbere y’igihe ukuntu Si igabanywa mu buryo butari bwo. Dufashe mu muzirikanyi ko hariho ibintu bimwebimwe bihindura ibiribwa biri mu bihe vyihariye mu bihe vyihariye, kandi ibindi bihimba ntibiri mu bihe vyihariye, kandi agaciro kavyo ko kwikingira kavana n’ibidukikije, twasavye kandi uburyo bwo kwikingira buvanze, aho δj na Hj bigabanywa mu bice bibiri ku ruhande rwa Si Ukwongerekana mu nzira zose, mu gihe imigwi imwe imwe gusa y’ibindi bihimba vy’umubiri izogira agaciro kanini k’ukwikingira, iyo Si ifise agaciro kanini k’ukwikingira. n’ugusangira (Ishusho 1B).
Kugira ngo tugerageze ubuhanuzi bw’inyigisho y’ukwikingira yasubiwemwo ku nzira y’insobanuro y’inyigisho y’amakuru, twareze utuvunji tw’ibikoko birya ivyatsi vy’abahinga (Ms) canke b’abahinga (Sl) ku mababi y’igiti citwa Nepenthes pallens (Ishusho 2A). Dukoresheje isesengura rya MS/MS, twaronse imirongo 599 y’imirongo ya MS/MS idasubira inyuma (dosiye y’amakuru S1) ivuye mu bihimba vy’amababi vy’ivyatsi vy’amababi vyakoranijwe inyuma yo kugaburira ibikoko. Gukoresha urutonde rwa RDPI, Hj, na δj kugira ngo umuntu abone mu ciyumviro ukuntu amakuru ahindurwa mu madosiye y’imiterere ya MS/MS birerekana uburyo bushimishije (Ishusho 2B). Ivyo muri rusangi bigenda ni uko, nk’uko bivugwa n’umusobanuzi w’amakuru, uko ibikoko bikomeza kurya amababi, urugero rw’ugusubira gutunganya ibintu vyose mu bijanye n’imibiri rugenda rurushirizaho uko igihe kigenda kirarenga: amasaha 72 inyuma y’aho iyo nyamaswa irya ivyatsi iriye, RDPI iraduga cane. Ugereranije n’ivyo gucungera bitagiramwo ivyago, Hj yaragabanutse cane, ivyo bikaba vyatewe n’uko urugero rw’ubuhinga bw’imibiri rwari rwarateye imbere, ivyo bikaba vyagereranywa n’urugero rwa δj. Ivyo bigaragara bihuye n’ivyo inyigisho ya OD ivuga, ariko ntibihuye n’ivyo inyigisho ya MT ivuga, iyo na yo ikaba yizera ko amahinduka ataco akora (atajanye n’inzira) mu rugero rw’ibinyabuzima bikoreshwa nk’uburyo bwo kwikingira (Ishusho 1B). Naho ibiri muri oral secretion (OS) elicitor content n’ingeso yo kugaburira ivyo bikoko bibiri birya ivyatsi bitandukanye, ukugaburira kwavyo gutaziguye kwatumye habaho amahinduka asa n’ayo mu bijanye n’inzira za Hj na δj mu kiringo c’amasaha 24 n’amasaha 72 y’iyimbura. Itandukaniro ryonyene ryabaye ku masaha 72 ya RDPI. Ugereranije n’ivyo vyatewe n’ukugaburira Ms, ivy’uguhindura ibiribwa muri rusangi vyatewe n’ukugaburira Sl vyari vyinshi.
(A) Igishushanyo c’igerageza: ingurube rusangi (S1) canke abahinga (Ms) birya ivyatsi bigaburirwa amababi y’ibimera vy’ibinogo, mu gihe ku bikoko birya ivyatsi vyigereranywa, OS ya Ms (W + OSMs) ikoreshwa mu gukorana n’ugutobora ibikomere vy’ibibanza vy’amababi bihuye. S1 (W + OSSl) ivyatsi canke amazi (W + W). Igicungera (C) ni ibabi ritarononekaye. (B) Ugushobora gutera (RDPI ugereranyije n’igicapo c’igenzura), ubudasa (index ya Hj) n’index y’ubuhinga (δj index) biharuwe ku ikarita idasanzwe y’ibinyabuzima (599 MS/MS; dosiye y’amakuru S1). Inyenyeri zigaragaza itandukaniro rikomeye hagati y’ibifungurwa vy’ibikoko birya ivyatsi bitaziguye n’umugwi w’abagenzuzi (Ikigeragezo ca t c’umunyeshure n’ikigeragezo ca t c’ababiri, *P<0.05 na ***P<0.001). ns, si ngombwa. (C) Igihe co gutorera umuti urutonde rw’ibintu nyamukuru (agasanduku k’ubururu, aside amino, aside y’ibinyabuzima n’isukari; dosiye y’amakuru S2) n’ivy’ihinduka ry’ibintu bidasanzwe (agasanduku gatukura 443 MS/MS; dosiye y’amakuru S1) inyuma y’ugutera ivyatsi. Ibara ry’umurongo ryerekeza ku 95% vy’igihe co kwizigira. Inyenyeri yerekana itandukaniro rikomeye hagati y'ubuvuzi n'ubugenzuzi [isesengura ry'uburyo bubiri (ANOVA), rikurikiwe n'itandukaniro rikomeye ry'ukuri (HSD) rya Tukey ku bijanye n'ukugereranya kwinshi kw'inyuma y'igihe, * P <0.05, ** P <0.01 Na *** P <0.001]. (D) Ukwihariza kw’ibibanza vy’ugushwiragira n’imirongo idasanzwe y’ibinyabuzima (ibipimo bisubirwamwo n’imiti itandukanye).
Kugira ngo tumenye nimba uguhinduka kw’ibihimba vy’umubiri biterwa n’ibikoko birya ivyatsi ku rugero rwa metabolome bigaragara mu mpinduka ziri ku rugero rw’ibihimba vy’umubiri vy’umuntu ku giti ciwe, twabanje kwibanda ku bihimba vy’umubiri vyari vyaratohojwe mbere mu mababi y’igiti citwa Nepenthes pallens gifise ubushobozi bwo guhangana n’ibikoko birya ivyatsi. Amide za phenolic ni ivyuma bifatanya hydroxycinnamamide-polyamine vyirundanira mu gihe udukoko turya ivyatsi kandi bizwi ko bigabanya ubushobozi bw’udukoko (32). Twararondeye ivy’imbere mu MS/MS bihuye maze ducapura imirongo yavyo y’imigenderanire (Ishusho S1). Ntibitangaje rero kubona ibintu biva kuri phenol bitagira uruhara rudasanzwe mu kwikingira ibikoko birya ivyatsi, nk’aside chlorogenic (CGA) na rutin, bigabanywa inyuma y’ukurya ivyatsi. Mu buryo butandukanye n’ubwo, ibikoko birya ivyatsi birashobora gutuma phenol amides zigira ubushobozi bwinshi cane. Ugukomeza kugaburira ivyo bikoko bibiri birya ivyatsi vyatumye habaho urutonde rw’ugukangura hafi rumwe rw’ibihimba vy’umubiri bita phenolamides, kandi ivyo vyari bigaragara canecane ku bijanye n’ugukora de novo kw’ibihimba vy’umubiri bita phenolamides. Ivyo nyene bizoboneka mu gihe co gutohoza inzira y’ibinyabuzima bita 17-hydroxygeranyl nonanediol diterpene glycosides (17-HGL-DTGs), ivyara umubare munini w’ibinyabuzima bita acyclic diterpenes bifise ubushobozi bwo kurwanya ibikoko birya ivyatsi (33), ivyo bikaba ari vyo Ms Feeding yabivuze n’ibinyabuzima bifise ubushobozi bwo kurwanya ibinyabuzima (33).
Intambamyi ishobora guterwa n’igerageza ry’ukugaburira ibikoko birya ivyatsi ataco bimaze ni itandukaniro riri hagati y’urugero rw’amababi n’igihe ibikoko birya ivyatsi birya, ivyo bikaba bituma bigorana gukuraho ingaruka zidasanzwe ziterwa n’ibikomere n’ibikoko birya ivyatsi. Kugira ngo dushobore gutorera umuti neza ubwoko bw’ibikoko birya ivyatsi bwihariye bw’inyishu y’ihinduka ry’amababi, twagereranyije ukugaburira utuvunji twa Ms na Sl mu gukoresha ubwo nyene OS (OSM na OSS1) zari ziherutse kwegeranywa ku rugero rwo gutera W rw’ibibanza vy’amababi bihuye. Ubwo buryo bwitwa ubuvuzi bwa W + OS, kandi buratuma ubuvuzi buja ku rugero rumwe mu gushinga igihe nyaco c’intango y’inyishu iterwa n’inyamaswa zirya ivyatsi ata ngaruka zitera urujijo z’itandukaniro ry’urugero canke ubwinshi bw’ugutakaza utugingo ngengabuzima (Ishusho 2A) (34). Dukoresheje umurongo w’isesengura n’uguharura wa MS/MS, twaronse imirongo 443 ya MS/MS (dosiye y’amakuru S1), yari ihuriye n’imirongo yari yarakoranijwe mbere ivuye mu ngeragezwa z’ibifungurwa bitaziguye. Isesengura ry’inyigisho y’amakuru y’iyi nkuru ya MS/MS ryerekanye ko gusubira gutegura porogarama y’ibihimba vy’umubiri vy’amababi mu kwigana ibikoko birya ivyatsi vyerekana ko hariho ivyiza vy’umubiri (Ishusho 2C). Cane cane, ugereranyije n’ubuvuzi bwa OSS1, OSM yatumye habaho ugutera imbere kw’ubuhinga bwa metabolome ku masaha 4. Birabereye kumenya ko ugereranije n’amakuru y’igerageza y’ukugaburira ibikoko birya ivyatsi, ubuhinga bwo guhindura ibiribwa buboneka mu kibanza c’ingero zibiri hakoreshejwe Hj na δj nk’ibiharuro be n’uburongozi bw’ubuhinga bw’ibiribwa mu kwishura ku buvuzi bw’ibikoko birya ivyatsi mu gihe c’igihe kigenda kirahinduka (Ishusho 2D). Muri ico gihe nyene, twagereranyije ibirimwo aside amino, aside organique n’isukari (dosiye y’amakuru S2) kugira ngo turondere nimba iyo ntumbero yo kwiyongera kw’ubuhinga bw’ivy’imibiri iterwa n’ugusubira gutunganya ivy’imibiri y’abantu mu kwishura ku bikoko birya ivyatsi vyiyumviwe ( Igishushanyo S2). Kugira ngo dushobore gusigura neza iyo nzira, twarakurikiranye kandi uburyo bwo kwirundanira kw’ibihimba vy’umubiri vy’inzira za phenolamide na 17-HGL-DTG twavuganye mbere. Ivyo bihimba vy’umubiri vy’ibikoko birya ivyatsi bihinduka bikaba uburyo bwo gusubira gutunganya ibintu mu buryo butandukanye mu guhindura ibiribwa vy’ibikoko (Ishusho S3). Ivyo bihimba vy’umubiri birimwo ibice vya coumarin na caffeoyl bikunda guterwa na OSS1, mu gihe OSMs zituma habaho uguterwa n’ibihimba vy’umubiri bita ferulyl. Ku bijanye n’inzira ya 17-HGL-DTG, OS itandukanye n’ibintu biva mu mazi y’i malonylation n’ivy’i dimalonylation vyabonetse (Ishusho S3).
Inyuma y’aho, twarize ubuhinga bwo kwandika buterwa na OS dukoresheje amakuru y’igihe, yigana gukoresha OSMs mu kuvura amababi y’amababi y’igiterwa c’uruzitiro mu bikoko birya ivyatsi. Ivyerekeye ubuhinga bwo gufata ivyerekanwa birahuye n’ubuhinga bwakoreshejwe muri iki cigwa c’ivy’imibiri (35). Ugereranije n’ugusubira gutunganya metabolome aho ubuhinga bwo guhindura ibiribwa bugenda burushirizaho cane cane uko igihe kigenda kirarenga, turabona ko amababi y’igihe gito ashobora gutera imbere, aho ubushobozi bwo kwandika (RDPI) n’ubuhinga (δj) biri ku 1 Hariho ukwiyongera gukomeye kw’amasaha, kandi ubudasa (Hj) bwagabanutse cane muri iki gihe c’imvugo ya BMPx, i y’ubuhinga bwo kwandika (Ishusho S4). Imiryango y’imitsi y’ibinyabuzima (nka P450, glycosyltransferase, na BAHD acyltransferase) iragira uruhara mu nzira yo gukoranya ibinyabuzima bidasanzwe biva mu bice vy’imiterere biva ku binyabuzima vy’intango, bikurikije uburyo bwa kera bwavuzwe haruguru bw’ubuhinga buhambaye. Nk’akarorero, inzira ya phenylalanine yarasuzumwe. Iryo sesangura ryemeje ko utugingo ngengabuzima nyamukuru mu guhindura ibiribwa bita phenolamide duterwa cane na OS mu bikoko birya ivyatsi ugereranyije n’ibimera bitakwegerewe, kandi ko bihuye cane mu buryo bigaragara. Igitera kwandika MYB8 n’imiterere y’imitsi PAL1, PAL2, C4H na 4CL biri mu ruzi rwo hejuru rw’iyi nzira vyerekanye ko kwandika gutangura kare. Acyltransferases zigira uruhara mu gukoranya kwa nyuma kwa phenolamide, nka AT1, DH29, na CV86, zigaragaza uburyo buramba bwo gutera imbere (Ishusho S4). Ivyo bimenyetso vyerekana ko ugutangura hakiri kare kw’ubuhinga bwo kwandika no gutera imbere mu nyuma kw’ubuhinga bwo guhindura ibiribwa ari uburyo bufatanye, bishobora kuba biterwa n’uburyo bwo gutegeka bujanye n’igihe butangura inyishu ikomeye yo kwikingira.
Ivyo bimenyetso vy’imisemburo y’ibimera bikora nk’igice kigenzura amakuru y’ibinyabuzima birya ivyatsi kugira ngo bisubire gutegura imiterere y’ibimera. Inyuma y’igereranyo ry’ibikoko birya ivyatsi, twagereranije ubushobozi bw’ibice vy’imitsi y’ibimera vy’ingenzi maze tubona ukuntu bifatanya hagati yavyo mu gihe c’igihe [Pearson correlation coefficient (PCC)> 0.4] (Ishusho 3A). Nk’uko vyari bitezwe, amahomoni y’ibimera ajanye n’ibinyabuzima arahuzwa mu nzira y’uguserura amahomoni y’ibimera. Ikindi, uburyo bwo guhindura ibiribwa (Si index) burashirwa kuri iyo nzira kugira ngo bugaragaze imitsi y’ibimera iterwa n’ubuvuzi butandukanye. Ivyiyumviro bibiri nyamukuru vy’inyishu yihariye y’ibikoko birya ivyatsi birakwegwa: kimwe kiri mu kigwi ca JA, aho JA (uburyo bwayo bukora mu buryo bw’ibinyabuzima JA-Ile) n’ibindi bikomoka kuri JA vyerekana amanota menshi cane ya Si; ikindi ni étilene (ET). Gibberellin yerekanye gusa ukwiyongera gusanzwe kw’ibinyabuzima birya ivyatsi, mu gihe izindi ntungamubiri z’ibimera, nka cytokinin, auxin, na aside abscisique, zari zifise ubushobozi buke bwo gutera ivyatsi. Ugereranije n’ugukoresha W + W gusa, ugukomeza agaciro k’isonga k’ibikomoka kuri JA biciye mu gukoresha OS (W + OS) birashobora guhindurwa mu bisanzwe bikaba ikimenyetso gikomeye c’ibikomoka kuri JA. Mu buryo butategerezwa, OSM na OSS1 zifise ibintu bitandukanye bitera elicitor zizwi ko zituma JA na JA-Ile zirundanira nk’ivyo. Mu buryo butandukanye n’ubwa OSS1, OSM ivyurwa cane kandi bikomeye na OSMs, mu gihe OSS1 idakomeza inyishu y’ibikomere vyo mu mutwe (Ishusho 3B).
(A) Isesengura ry’uruja n’uruza rw’ibimenyetso bishingiye ku kubara kwa PCC kw’ivyigwa vy’imitsi y’ibimera biterwa n’ibikoko birya ivyatsi. Ico gice c’umubiri kigereranya umusemburo umwe w’igiterwa, kandi ubunini bw’ico gice c’umubiri bugereranya urutonde rwa Si rwihariye ku musemburo w’igiterwa hagati y’imiti. (B) Ukwirundanira kwa JA, JA-Ile na ET mu mababi bitewe n’imiti itandukanye yerekanwa n’amabara atandukanye: apricot, W + OSM; ubururu, W + OSSl; umukara, W + W; imvi, C (ugucungera) . Inyenyeri zigaragaza itandukaniro rikomeye hagati y’ubuvuzi n’ubugenzuzi (ANOVA y’inzira zibiri ikurikiwe n’ukugereranya kenshi kwa Tukey HSD post hoc, *** P <0.001). Isesengura ry’inyigisho y’amakuru ya (C)697 MS/MS (dosiye y’amakuru S1) mu bihimba vy’umubiri vya JA n’ubumuga bwo gutahura (irAOC na irCOI1) na (D)585 MS/MS (dosiye y’amakuru S1) muri ETR1 n’ikimenyetso ca ET gihungabanye Umugambi wo kugenzura umurongo w’ibikoko birya ivyatsi (umugambi wo gucungera ibikoko birya ivyatsi) wavyuriwe. Inyenyeri zigaragaza itandukaniro rikomeye hagati y’ubuvuzi bwa W+OS n’ubugenzuzi butagira ico bukoze (ANOVA y’inzira zibiri ikurikiwe n’ukugereranya kenshi kwa Tukey HSD, *P<0.05, **P<0.01 na ***P<0.001). (E) Ivyerekana binyanyagiye vy’ukurwanya gunyanyagiye kw’ubuhinga bwihariye. Amabara agereranya imiryango itandukanye yahinduwe mu vy’uruvyaro; ibimenyetso bigereranya uburyo butandukanye bwo kuvura: inkomoko y’inkomoko, W + OSS1; umurongo w’inzira, W + OSM; uruziga C
Inyuma y’aho, dukoresha ubwoko bwa Nepenthes buhinduwe mu buryo bw’ivy’ivyaduka (irCOI1 na sETR1) mu ntambwe nyamukuru z’ubuhinga bwa JA na ET (irAOC na irACO) n’ugutahura (irCOI1 na sETR1) kugira ngo dusuzume ukuntu izo nzira zibiri z’ibimera zihinduka ku ruhara rw’ibikoko birya ivyatsi. Mu buryo buhuye n’ivyageragejwe vya kera, twemeje ko herbivore-OS yinjijwe mu bimera bifise ubusa (EV) (Ishusho 3, C gushika kuri D) be n’ukugabanuka muri rusangi kw’urugero rwa Hj rwatewe na OSM, mu gihe urugero rwa δj rwariyongereye. Inyishu iragaragara cane kuruta inyishu yavyurwa na OSS1. Igishushanyo c’imirongo ibiri gikoresha Hj na δj nk’ibiharuro vyerekana ukuntu ivy’ugukuraho amategeko vyihariye (Ishusho 3E). Ico kigaragara cane ni uko mu mice idafise ikimenyetso ca JA, ubudasa bw’imibiri n’ihinduka ry’ubuhinga ry’ibinyabuzima biterwa n’ibikoko birya ivyatsi hafi bikurwaho (Ishusho 3C). Mu buryo bunyuranye, ukuntu ET igaragara mu bimera vya sETR1, naho ingaruka rusangi ku mahinduka mu mibiri y’ibikoko birya ivyatsi iri hasi cane y’iy’ibimenyetso vya JA, biragabanya itandukaniro ry’ibimenyetso vya Hj na δj hagati y’ibivyuka vya OSM na OSS1 (Ishusho 3D n’Ishusho S5). . Ivyo vyerekana ko uretse igikorwa nyamukuru c’uguhindura ikimenyetso ca JA, uguhindura ikimenyetso ca ET na co nyene gikora nk’ugutunganya neza inyishu y’ubwoko bw’ibikoko birya ivyatsi. Mu buryo buhuye n’ico gikorwa co gutunganya neza, nta mpinduka yabaye mu bijanye n’uguhindura ibihimba vy’umubiri mu bimera vya sETR1. Ku rundi ruhande, ugereranyije n’ibiterwa vya sETR1, ibiterwa vya irACO vyatumye habaho amahinduka asa n’ayo mu bijanye n’imibiri bitewe n’ibikoko birya ivyatsi, ariko vyerekana ko hariho ibiharuro bitandukanye cane vya Hj na δj hagati y’ingorane za OSM na OSS1 (Ishusho S5).
Kugira ngo tumenye ibintu bidasanzwe bihindura ibiribwa bifise uruhara runini mu kwishura kw’ubwoko bw’ibikoko birya ivyatsi no gutunganya neza ivyimburwa vyavyo biciye ku bimenyetso vya ET, twakoresheje uburyo bwa MS/MS bwari bwateguwe mbere. Ubu buryo bushingiye ku buryo bwo gukoranya ibice bibiri kugira ngo umuntu asubire guca irya n’ino umuryango w’ibinyabuzima biva mu bice vya MS/MS [normalized dot product (NDP)] n’igiharuro c’ugusa gishingiye ku gutakaza ataco bimaze (NL). Ivyiyumviro vya MS/MS vyubatswe biciye mu gusesangura imirongo ya ET transgenic vyavuyemwo 585 MS/MS (dosiye y'amakuru S1), ivyo bikaba vyatowe umuti mu kubishira mu bice indwi vy'ingenzi vya MS/MS (M) (Ishusho 4A) . Bimwe muri ivyo bihimba vyuzuyemwo ibintu bidasanzwe vyari vyaramenyekanye mbere: nk’akarorero, M1, M2, M3, M4 na M7 birimwo ibintu bitandukanye biva ku bihimba vy’umubiri (M1), ibihimba vy’umubiri bita flavonoïdes (M2), isukari y’ibihimba vy’umubiri (M3 Na M4), na 17-HGL-DTG (M7). Ikindi, amakuru yihariye yerekeye ivy’imibiri (Si index) y’imibiri imwe muri buri kiganiro araharurirwa, kandi ukuntu Si igabanywa bishobora kubonwa mu buryo busanzwe. Muri make, imirongo ya MS/MS yerekana ivyatsi vyinshi n’ubudasa bw’ubwoko bw’umubiri irangwa n’agaciro kanini ka Si, kandi imibare ya kurtosis yerekana ukuntu ubwoya bukwiragira ku mfuruka y’umurizo w’iburyo. Imwe muri izo nzira z’ugusangira kw’ibihimba vy’umubiri bitagira ibinure vyabonetse muri M1, aho phenol amide yerekanye igice ca Si kinini kuruta ibindi vyose (Ishusho 4B). 17-HGL-DTG y’ibikoko birya ivyatsi twavuze mbere muri M7 yerekanye igiharuro ca Si kiringaniye, kigaragaza urugero ruringaniye rw’ugutunganya itandukaniro hagati y’ubwoko bubiri bwa OS. Mu buryo bunyuranye, vyinshi mu bihimba vy’umubiri bikora mu buryo bwihariye, nka rutin, CGA, n’isukari ya acyl, biri mu bihimba vy’umubiri bifise amanota make cane ya Si. Kugira ngo haboneke neza uburemere bw’imiterere n’ugusangira kwa Si hagati y’ibinyabuzima bidasanzwe, hakozwe uruja n’uruza rw’ibihimba vy’umubiri ku gice kimwekimwe cose (Ishusho 4B). Iciyumviro gihambaye c’ivyiyumviro vya OD (bivugwa mu ncamake mu gishushanyo 1B) ni uko ugusubira gutunganya ibintu bidasanzwe bihindura ibiribwa inyuma y’ukurya ivyatsi bikwiye gutuma habaho amahinduka y’inzira imwe mu bihimba vy’umubiri bifise agaciro kanini ko kwikingira, cane cane mu kwongera ubudasa bwavyo (bitandukanye n’ugukwiragira kw’imburakimazi) Uburyo) Ivyiyumviro vy’ukwikingira vyavuzwe mbere n’ivyiyumviro vya MT. Ivyinshi mu biva kuri phenol vyirundanijwe muri M1 birafitaniye isano n’ugugabanya ubushobozi bw’udukoko (32). Igihe twagereranya agaciro ka Si mu bihimba vy’umubiri vya M1 hagati y’amababi yandujwe n’amababi agize ibiterwa bigenzura EV ku masaha 24, twabonye ko ubudasa bw’ibihimba vy’umubiri vy’ibihimba vyinshi vy’umubiri inyuma y’udukoko birya ivyatsi bufise umurongo ukomeye wo kwiyongera (Ishusho 4C). Ukwo kwongerekana kw’agaciro ka Si kwabonetse gusa mu bihimba vy’umubiri bikingira, ariko nta kwongerekana kw’agaciro ka Si kwabonetse mu bindi bihimba vy’umubiri bita phenols n’ibindi bihimba bitazwi bibaho muri iki kiganiro. Ico ni ikigereranyo kidasanzwe, gifitaniye isano n’inyigisho ya OD. Ivyiyumviro nyamukuru vy’ihinduka ry’imibiri biterwa n’ibikoko birya ivyatsi birahuye. Kugira ngo tugerageze nimba iyo nzira yihariye y’uruzitiro rwa phenolamide yatumwe na ET yihariye ya OS, twaciye ducapura urutonde rw’ibinyabuzima Si maze dutuma haba agaciro k’imvugo itandukanye hagati ya OSM na OSS1 mu genotypes za EV na sETR1 (Ishusho 4D ). Mu sETR1, itandukaniro ry’imiti yitwa phenamide hagati ya OSM na OSS1 ryaragabanutse cane. Uburyo bwo gukoranya ibice bibiri bwakoreshejwe kandi ku makuru ya MS/MS yashizwe hamwe mu mice ifise JA idahagije kugira ngo haboneke ibice nyamukuru vya MS/MS bijanye n’ubuhinga bwo guhindura ibiribwa bugenwa na JA (Ishusho S6).
(A) Ivyavuye mu gukoranya 585 MS/MS bishingiye ku gice gisangiye (ugusa kwa NDP) n’ugutakaza ataco bihuriyeko (ugusa kwa NL) bituma igice (M) gihuye n’umuryango w’ibirungo bizwi, canke n’ibirungo vy’ibirungo bitazwi canke bihinduwe nabi. Inyuma y’igice kimwekimwe cose, harerekanwa ukuntu ibintu bihinduka (MS/MS) vyihariye (Si) vyerekanywe. (B) Urusobe rw’ibihimba vy’umubiri: Ivyuma bigereranya MS/MS n’imirongo, NDP (umutuku) na NL (ubururu) MS/MS (cut-off,> 0.6). Igiharuro c’ubudasa bw’ibinyabuzima (Si) gifise ibara rishingiye ku gice (ibubamfu) kandi kigashirwa ku ikarita y’uruzitiro rw’ibinyabuzima (iburyo). (C) Module M1 y’igiterwa ca EV mu gihe c’ubuhinga (ubugenzuzi) n’ubuhinga (ikigereranyo c’ibikoko birya ivyatsi) ku masaha 24: igishushanyo c’uruja n’uruza rw’ibihimba (agaciro ka Si ni ubunini bw’urudodo, phenolamide y’ukwikingira iragaragazwa n’ubururu). (D) Igishushanyo c’uruzitiro rw’amamolekule M1 c’umurongo w’imirongo sETR1 n’ugutahura kwa EV na ET guhungabanye: ikirungo ca phenolic kigereranywa n’uruziga rw’icatsi kibisi, n’itandukaniro rikomeye (agaciro ka P) hagati y’imiti ya W + OSM na W + OSS1 nk’ubunini bw’uruziga. CP, N-kafeyoyili-tirozine; CS, N-kafeyoyili-ispermidine; FP, aside N-ferulique y’ester-urique; FS, N-ferulili-siperimidini; CoP, N’, N “-Komaroli-tirozine; DCS, N’, N”-dikafeoyil-spermidine; CFS, N’, N”-kafeyoyili, feruloyili-spermidine;
Twarabandanije kwagura isesengura kuva ku genotype imwe ya Nepenthes yagabanutse gushika ku bantu b’ibinyabuzima, aho amahinduka akomeye hagati y’ubwoko bw’ibikoko birya ivyatsi mu rugero rwa JA n’urugero rw’ibinyabuzima vyihariye vyari vyaradondowe mbere mu bantu b’ibinyabuzima (26). Koresha aya makuru kugira ngo upfuke imigera 43. Ivyo bihimba vy’umubiri birimwo ubwoko 123 bw’ibimera biva mu giti citwa N. pallens. Ivyo bitegwa vyakuwe mu mbuto zakwegeranijwe mu bibanza bitandukanye vy’imvukira muri Utah, Nevada, Arizona, na California (Ishusho S7), twaharuriye ubudasa bw’ibinyabuzima (aha bwitwa urugero rw’abantu) ubudasa bwa β) n’ubudasa bw’ibinyabuzima buterwa na OSM. Mu buryo buhuye n’ivyigwa vya kera, twabonye amahinduka menshi y’imibiri ku nzira ya Hj na δj, ivyo bikaba vyerekana ko imigera ifise itandukaniro rikomeye mu bijanye n’ubuhinga bwo guhindura ibiri mu mubiri ku bikoko birya ivyatsi (Ishusho S7). Iryo shirahamwe ryibutsa ivyo twabonye mbere ku bijanye n’ihinduka ry’inguvu ry’ibinyabuzima bita JAs riterwa n’ibikoko birya ivyatsi, kandi ryagumye rifise agaciro kanini cane mu bantu bamwe (26, 36). Mu gukoresha JA na JA-Ile kugira ngo tugerageze isano ry’urugero rusangi hagati ya Hj na δj, twabonye ko hariho isano ryiza rikomeye hagati ya JA n’ubudasa bw’ibinyabuzima β n’urutonde rw’ubuhinga (Ishusho S7). Ivyo vyerekana ko ubudasa buterwa n’ibikoko birya ivyatsi mu gutera JAs buboneka ku rugero rw’abantu bushobora kuba buterwa n’uguhinduka kw’ibihimba vy’umubiri bitewe n’uguhitamwo mu bikoko birya ivyatsi vy’udukoko.
Ivyigwa vya kera vyerekanye ko ubwoko bw’itabi butandukanye cane mu bwoko no mu bijanye n’ukuntu buvana n’uburyo bwo kwikingira umubiri. Bivugwa ko ayo mahinduka mu guhindura ikimenyetso c’ibikoko birya ivyatsi n’ubushobozi bwo kwikingira bigenwa n’inguvu z’udukoko, ubuzima bw’ibimera, n’ibiciro vyo kwikingira mu kibanza aho ubwoko bumwe bumwe bukura. Twarasuzumye ukuntu amababi ahinduka mu buryo buhuye n’ibikoko birya ivyatsi vy’ubwoko butandatu bw’ibikoko vyitwa Nicotiana bikomoka muri Amerika y’Uburaruko na Amerika y’Epfo. Ubwo bwoko burafitaniye isano cane n’igiti citwa Nepenthes muri Amerika y’Uburaruko, ari co Nicolas Bociflo. La, N. nikotini, Nikoti n. ivyatsi bigoyagoya, itabi ry’umurongo, itabi ry’umukara (Nicotiana spegazzinii) n’itabi ry’amababi y’itabi (Ishusho 5A) (37). Ivyatsi bitandatu muri ivyo, harimwo n’ubwoko buzwi neza N. please, ni ibimera bimera umwaka wose vyo mu bwoko bwa petunia, obtusifolia N. na yo ni ibimera bihoraho vyo mu bwoko bwa mushiki wacu Trigonophyllae (38). Hanyuma, W + W, W + OSM na W + OSS1 induction vyakozwe kuri izo nyamaswa indwi kugira ngo higwe ukuntu ubwoko busubira gutunganya ivy’ukurya kw’udukoko.
(A) Igiti c’ubuhinga bwa bootstrap gishingiye ku vyo bishoboka cane [ku bijanye n’ugukora glutamine y’umubiri (38)] be n’ugukwiragira kw’ubwoko indwi bwa Nicotiana bufitaniye isano rya hafi (amabara atandukanye) (37). (B) Igishushanyo c’ugushwiragira kw’ubudasa bwihariye bw’imiterere y’ibifungurwa vy’ubwoko indwi bwa Nicotiana (939 MS/MS; dosiye y’amakuru S1). Ku rugero rw’ubwoko, ubudasa bw’ibinyabuzima burafitaniye isano ribi n’urugero rw’ubuhinga. Isesengura ry’isano hagati y’ubudasa bw’ibifungurwa n’ubuhinga n’ukwirundanira kwa JA vyerekanywe ku gishushanyo ca 2. S9. Ibara, ubwoko butandukanye; inkomoko y’inkomoko, W + OSS1; umurongo w’inzira, W + OSM; (C) Nicotiana JA na JA-Ile zishirwa ku rutonde hakurikijwe uburemere bw’ugukangura kwa OS (ANOVA y’inzira zibiri na Tukey HSD inyuma y’ukugereranya kwinshi, * P <0.05, ** P <0.01 na * ** Kugereranya W + OS na W + W, P <0.001). Igishushanyo c’agasanduku k’ubudasa (D) n’ubudasa bw’ubwoko bumwe bumwe inyuma yo kwigana ibikoko birya ivyatsi n’ivyo bita methyl JA (MeJA). Inyenyeri yerekana itandukaniro rikomeye hagati ya W + OS na W + W canke lanolin yongeweko W (Lan + W) canke Lan yongeweko MeJA (Lan + MeJa) n’ubugenzuzi bwa Lan (isesengura ry’inzira zibiri ry’ugutandukanya, hakurikijwe ukugereranya kw’incuro nyinshi kwa Tukey kwa HSD , *P<0.05, **P<0.01 na).
Dukoresheje uburyo bw’ibice bibiri, twabonye ibice 9 vy’ibice 939 MS/MS (dosiye y’amakuru S1). Ivyiyumviro vya MS/MS vyasubiwemwo n’imiti itandukanye biratandukanye cane hagati y’ibice bitandukanye hagati y’ubwoko (Ishusho S8). Guca mu ciyumviro Hj (ivugwa aha nk’ubwoko bw’ubwoko γ-ubudasa) na δj birerekana ko ubwoko butandukanye buteranira hamwe mu migwi itandukanye cane mu kibanza c’imibiri, aho ugucapura ku rugero rw’ubwoko akenshi kugaragara cane kuruta ugukangura. Uretse N. linear na N. obliquus, zigaragaza ingaruka nyinshi z’inguvu z’ugutera (Ishusho 5B). Mu buryo bunyuranye, ubwoko nka N. purpurea na N. obtusifolia burafise inyishu igaragara cane y’imibiri iyo buvuwe, ariko metabolome iratandukanye cane. Ugukwiragizwa kw’ubwoko bwihariye kw’inyishu y’imibiri yatumye habaho isano ribi hagati y’ubuhinga n’ubudasa bwa gamma (PCC = -0.46, P = 4.9×10-8). Impinduka ziterwa na OS mu rwego rwa JAs zifitaniye isano n’ubuhinga bw’imibiri, kandi zifitaniye isano ribi n’ubudasa bw’imibiri bugaragazwa n’ubwoko bumwe bumwe (Ishusho 5B n’Ishusho S9). Birabereye kumenya ko ubwoko buvugwa mu rurimi rw’ikirundi ko ari ubwoko bw’“inyishu y’ikimenyetso” ku gishushanyo ca 5C, nk’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko, ibikoko vy’ibikoko vy’ibikoko, ibikoko vy’ibikoko vy’ibikoko vy’ibikoko, n’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko, vyateye ibimenyetso bihambaye ku minota 30. Ivyimburwa vya JA na JA-Ile biherutse gutera, mu gihe izindi migera zitwa “ikimenyetso kitagira ico zikora”, nk’insyo za Nepenthes, Nepenthes powdery na N. obtusifolia zigaragaza gusa induction ya JA-Ile Edge ata OS yihariye (Ishusho 5C). Ku rugero rw’imibiri, nk’uko twabibonye haruguru, ku Nepenthes zicitse intege, ibintu vyishura ku kimenyetso vyerekana ko OS idasanzwe kandi vyiyongera cane δj, mu gihe bigabanya Hj. Ivyo bimenyetso vy’ugutangura bishingiye kuri OS ntivyabonetse mu bwoko bwashizwe mu rwego rw’ubwoko butagira ikimenyetso (Ishusho 5, D na E). OS-specific metabolites zisangizwa kenshi hagati y’ubwoko bushobora kwishura ku bimenyetso, kandi ayo matsinda y’ibimenyetso akorana n’ubwoko bufise inyishu z’ibimenyetso zigoyagoya, mu gihe ubwoko bufise inyishu z’ikimenyetso zigoyagoya bugaragaza ko bufitaniye isano ritoyi (Ishusho S8 ). Ico ciyumviro kigaragaza ko OS-specific induction ya JAs n’ugusubira gutunganya OS-specific metabolome y’inyuma y’uruzi bifatanye ku rugero rw’ubwoko.
Inyuma y’aho, twakoresheje umuti wa lanolin urimwo methyl JA (MeJA) kugira ngo tuvure ibitegwa kugira ngo turondere nimba izo nzira zo gufatanya zibujijwe n’ukuboneka kwa JA ikoreshwa na JA y’inyuma, izoba iri muri cytoplasme y’ibitegwa. Gukuraho ivyuma vyihuta ni JA. Twabonye iyo nzira nyene y’uguhinduka buhoro buhoro kuva ku bwoko bushobora gusubiza ikimenyetso gushika ku bwoko butasubiza ikimenyetso bitewe n’ugukomeza gutanga JA (Ishusho 5, D na E). Muri make, ubuvuzi bwa MeJA bwahinduye cane porogarama y’ibihimba vy’umubiri vy’ibikoko bifise umurongo, N. obliquus, N. aquaticus, N. pallens, na N. mikimotoi, bituma δj yiyongera cane, Hj na yo igabanuka. N. purpurea yerekanye gusa ko δj yiyongera, ariko si Hj. N. obtusifolia, mbere yerekanwe ko yirundanira ivyiyumviro bike cane vy’ama JA, na yo nyene ntiyishura neza ku buvuzi bwa MeJA mu bijanye no gusubira gutunganya ivyuma bita metabolome. Ivyo bimenyetso vyerekana ko uguhingura JA canke uguhindura ikimenyetso bibujijwe mu buryo bw’umubiri mu bwoko butagira ikimenyetso. Kugira ngo tugerageze iyo nkuru, twarize ubwoko bune (N. pallens, N. mills, N. pink na N. microphylla) buterwa n’ibinyabuzima bita W + W, W + OSMs na W + OSS1 (39). Mu buryo buhuye n’ingene metabolome ihindura, ubwoko buratandukanye neza mu kibanza c’inyandiko, muri vyo N. attenuated yerekanye RDPI iterwa na OS ihambaye cane, mu gihe N. gracilis yari ifise RDPI iri hasi cane (Ishusho 6A). Ariko rero, vyabonetse ko ubwoko bw’ibihimba vy’umubiri buterwa na N. oblonga ari bwo buri hasi cane mu bwoko bune, bitandukanye n’ubwoko bwinshi bw’ibihimba vy’umubiri bwa N. oblonga bwari bwagaragajwe mbere mu bwoko indwi. Ivyigwa vya kera vyerekanye ko umugwi w’imitsi y’umubiri ijyanye n’ibimenyetso vy’ukwikingira vya kera, harimwo n’ibimenyetso vya JA, bisigura ubudasa bw’inyishu z’ukwikingira za kera ziterwa n’ibinyabuzima bifitaniye isano n’ibikoko birya ivyatsi mu bwoko bwa Nicotiana (39). Kugereranya inzira z’ibimenyetso vya JA hagati y’izo nyamaswa zine vyagaragaje uburyo bushimishije (Ishusho 6B). Ivyiyumviro vyinshi biri muri iyo nzira, nka AOC, OPR3, ACX na COI1, vyerekanye urugero rwo hejuru rw’ugutera imbere muri izo nzira zine. Ariko rero, umugera w’ingenzi, JAR4, uhindura JA mu buryo bwayo bukora mu buryo bw’ibinyabuzima bw’ivyanditswe vyirundanijwe na JA-Ile, kandi urugero rw’ivyanditswe vyawo ruri hasi cane, cane cane muri N. mills, Nepenthes pieris na N. microphylla. Ikindi, ni inyandiko y’iyindi gene AOS gusa itabonetse muri N. bifidum. Ivyo bihinduka mu guserura ivyiyumviro vy’umubiri bishobora kuba ari vyo bituma habaho ivyiyumviro bikomeye cane biterwa n’uguhingura JA muto mu bwoko bw’ibimenyetso bitagira ubushobozi bwo gukora be n’uguhingura N. gracilis.
(A) Isesengura ry’inyigisho y’amakuru y’ugusubira gutegura inyishu z’inyandiko za kera z’ubwoko bune bw’itabi bufitaniye isano rya hafi bwafashwe nk’akarorero iminota 30 inyuma y’ugutera ivyatsi. RDPI ibarwa mu kugereranya amababi aterwa n’ibikoko birya ivyatsi OS n’ivyo gucungera ibikomere. Amabara yerekana ubwoko butandukanye, ibimenyetso na vyo vyerekana uburyo butandukanye bwo kuvura. (B) Isesengura ry’imiterere y’imitsi mu nzira z’ibimenyetso vya JA mu bwoko bune. Inzira ya JA yoroshe yerekanwa iruhande y'igishushanyo c'agasanduku. Amabara atandukanye yerekana uburyo butandukanye bwo gutunganya. Inyenyeri yerekana ko hariho itandukaniro rikomeye hagati y’ubuvuzi bwa W + OS n’ubuvuzi bwa W + W (ku bijanye n’ikigeragezo ca t c’Umunyeshure ku bijanye n’itandukaniro ry’ababiri babiri, *P<0.05, **P<0.01 na ***P<0.001). OPDA, aside y’ibimera 12; OPC-8: 0,3-oxo-2(2′(Z)-pentenili)-sikolopentani-1-aside y’oktanoyike.
Mu gice ca nyuma, twarize ingene uguhindura ubwoko bw’udukoko mu bijanye n’imibiri y’ubwoko butandukanye bw’ibimera bishobora guhangana n’ibikoko birya ivyatsi. Ubushakashatsi bwa kera bwashimikiye cane ku bwoko bwa Nicotiana. Ubushobozi bwavyo bwo kurwanya Ms n’ibikoko vy’ibitoke buratandukanye cane (40). Aha, twarize isano riri hagati y’ico kigereranyo n’ubuhinga bwabo bwo guhindura ibiribwa. Dukoresheje ubwoko bune bw’itabi bwavuzwe haruguru, no kugerageza isano hagati y’ubudasa n’ubudasa bw’ibinyabuzima biterwa n’ibikoko birya ivyatsi n’ubushobozi bw’ibimera bwo kurwanya Ms na Sl, twabonye ubudasa, ubudasa n’ubudasa bw’ibinyabuzima ku muganga rusangi Sl Vyose bifitaniye isano ryiza, mu gihe isano hagati y’ubudasa bw’abagore b’abahinga ari, n’ S10). Ku bijanye n’ubushobozi bwo guhangana na S1, N. chinensis na N. gracilis zompi zacitse intege, zari zarerekanwe mbere ko zigaragaza ingero zompi z’uguhindura ikimenyetso ca JA n’ubuhinga bwo guhindura ibiri mu mubiri, zari zifise inyishu zitandukanye cane ku bijanye n’ugutera imbere kw’ibikoko birya ivyatsi, kandi na zo nyene zari zifise ubushobozi bwo guhangana n’ibikoko birya ivyatsi. Ivy'igitsina.
Mu myaka mirongo itandatu iheze, inyigisho y’ukwigwanira kw’ibimera yaratanze urutonde rw’ivyiyumviro, abashakashatsi bashingiye kuri vyo bavuze imbere y’igihe umubare munini w’iterambere n’ibikorwa vy’ibimera vyihariye bihindura ibiribwa. Vyinshi muri ivyo vyiyumviro ntibikurikiza uburyo busanzwe bwo gufata ingingo zikomeye (41). Batanga ubuhanuzi nyamukuru (3) ku rugero rumwe rw’isesengura. Igihe igerageza ry’ibivugwa imbere y’igihe ry’ingenzi rituma ivyiyumviro vyihariye bisuzumwa, ivyo bizotuma The field is moving forward. Mushigikirwe, ariko mukanke abandi (42). Ahubwo, iyo nyigisho nshasha iratanga ubuhanuzi ku nzego zitandukanye z’isesengura kandi yongerako urugero rushasha rw’ibintu bidondora (42). Ariko rero, izo nyigisho zibiri zashikirijwe ku rugero rw’ibikorwa, MT na OD, zirashobora gusigurwa mu buryo bworoshe nk’ubuhanuzi buhambaye bw’amahinduka yihariye y’imibiri aterwa n’ibikoko birya ivyatsi: inyigisho ya OD yizera ko amahinduka mu “kibanza” kidasanzwe c’imibiri ari ay’inzira nyinshi . Ivyiyumviro vya MT vyizera ko ayo mahinduka azoba ata nzira kandi azoba ari mu kibanza c’ibiribwa, kandi akunda kugira ibiribwa bifise agaciro kanini ko kwikingira. Ivyigwa vya kera vy’ubuhanuzi bwa OD na MT vyageragejwe hakoreshejwe umugwi mutoyi w’ibintu vy’“ukwikingira” vy’imbere. Ivyo bipimo bishingiye ku bihimba vy’umubiri bibuza ubushobozi bwo gusuzuma urugero n’inzira y’ugusubira gutunganya ibihimba vy’umubiri mu gihe c’ukurya ivyatsi, kandi ntivyemerera gupima mu rwego rw’imibare rudahinduka kugira ngo bisabe ubuhanuzi nyamukuru bushobora gufatwa nk’uburyo bwose Gupima amahinduka mu bihimba vy’umubiri vy’ibimera. Aha, twakoresheje ubuhinga bushasha mu vy’ubuhinga bw’imibiri bushingiye ku buhinga bwa MS bushingiye ku buhinga bwa none kandi dukora isesengura ry’ubuhinga bwa deconvolution MS mu mafaranga rusangi y’ibisobanuro vy’inyigisho z’amakuru kugira ngo tugerageze itandukaniro hagati y’ivyo bibiri vyasabwe ku rugero rw’ubuhinga bw’imibiri y’abantu kw’isi yose. Iciyumviro nyamukuru c’ubuhanuzi bw’iyi nyigisho. Ivyiyumviro vy’amakuru vyakoreshejwe mu bintu vyinshi cane cane mu bijanye n’ubushakashatsi ku bijanye n’ubudasa bw’ibinyabuzima n’ugutemba kw’ingaburo (43). Ariko rero, nk’uko tubizi, iyi ni yo nzira ya mbere ikoreshwa mu kudondora ikibanza c’amakuru y’ibinyabuzima no gutorera umuti ingorane z’ibidukikije zijanye n’ihinduka ry’ibinyabuzima ry’igihe gito mu kwishura ku bimenyetso vy’ibidukikije. Cane cane, ubushobozi bw’ubu buryo buri mu bushobozi bwabwo bwo kugereranya uburyo buri hagati y’ubwoko bw’ibimera no hagati yavyo kugira ngo hasuzumwe ingene ibikoko birya ivyatsi vyavuye mu bwoko butandukanye bikaja ku buryo bukomeye hagati y’ubwoko ku rwego rw’ubwihindurize butandukanye. Uguhindura ibiribwa.
Isesengura ry’ibice nyamukuru (PCA) rihindura amakuru menshi mu kibanza co kugabanya uburebure kugira ngo umurongo nyamukuru w’amakuru ushobore gusigurwa, rero akenshi rikoreshwa nk’ubuhinga bwo gutohoza kugira ngo umuntu asesengure amakuru, nk’ivyo bita deconvolution metabolome. Ariko rero, kugabanya uburebure bizotakaza igice c’amakuru ari mu rutonde rw’amakuru, kandi PCA ntishobora gutanga amakuru y’ingero yerekeye ibiranga bifitaniye isano cane n’inyigisho y’ibidukikije, nk’akarorero: ingene ibikoko birya ivyatsi bisubira gutunganya ubudasa mu bikorwa vyihariye (nk’akarorero, ubutunzi, ukwiragira) N’ubwinshi) ibintu bihindura ibiribwa? Ni ibihe bihimba vy’umubiri bishobora gutuma umuntu agira ingorane y’inyamaswa irya ivyatsi? Uvuye ku bijanye n’ubudasa, ubudasa n’ubushobozi bwo gutera, amakuru ari mu bijanye n’imiterere y’ibinyabuzima vy’amababi arabora, kandi bigaragara ko kurya ibikoko birya ivyatsi bishobora gutuma habaho ihinduka ry’ibinyabuzima. Mu buryo tutavyiteze, twabonye ko, nk’uko vyadondowe mu bimenyetso vy’inyigisho y’amakuru vyashizwe mu ngiro, ivy’uguhindura ibifungurwa biva muri ivyo bifise uguhuza gukomeye inyuma y’ibitero vy’ivyo bikoko bibiri birya ivyatsi (umunyagihugu Sl agaburirwa ijoro) n’umuhinga mu vy’ibimera vy’ibitoke vyitwa Solanaceae Ms. Igikoresho co gutanguza aside y’amavuta n’aside y’amavuta (FAC) muri OS (31). Mu gukoresha OS y’ibikoko birya ivyatsi mu kuvura ibikomere vy’ugutobora bihuye n’ingingo ngenderwako, ubuvuzi bw’ibikoko birya ivyatsi bwigana na bwo nyene bwarerekanye ingendo nk’iyo. Ubwo buryo busanzwe bwo kwigana ingene ibimera vyishura ku bitero vy’ibikoko birya ivyatsi burakuraho ibintu bitera urujijo bitewe n’uguhinduka kw’inyifato yo kurya y’ibikoko birya ivyatsi, ivyo bikaba bituma habaho ivyago bitandukanye mu bihe bitandukanye (34). FAC, izwi ko ari yo itera cane OSM, igabanya JAS n’izindi nyishu z’imitsi y’ibimera muri OSS1, mu gihe OSS1 igabanya incuro amajana (31). Ariko rero, OSS1 yatumye habaho ingero zisa n’izo z’ukwirundanira kwa JA ugereranyije na OSM. Vyaragaragajwe mbere ko inyishu ya JA mu Nepenthes yacitse intege igira ico ikoze cane ku OSM, aho FAC ishobora kuguma ikora naho yoba yashizwemwo amazi 1:1000 (44). Rero, ugereranyije na OSM, naho FAC muri OSS1 iri hasi cane, irahagije kugira ngo ivyure ikiza ca JA gihagije. Ivyigwa vya kera vyerekanye ko poroteyine zisa n’iz’ibinyabuzima (45) n’ibinyabuzima bita oligosaccharides (46) bishobora gukoreshwa nk’ibimenyetso vy’ibihimba vy’umubiri kugira ngo bivyure inyishu z’ukwikingira kw’ibimera muri OSS1. Ariko rero, ntibiramenyekana nimba izo elicitors ziri muri OSS1 ari zo zituma JA yirundanira nk’uko vyabonetse muri iki cigwa.
Naho hariho inyigisho nkeyi zidondora ibimenyetso vy’intoke bitandukanye vy’imibiri biterwa n’ugukoresha ibikoko bitandukanye birya ivyatsi canke JA canke SA (acide salicylique) (47), nta n’umwe yahungabanije ubwoko bw’ibikoko birya ivyatsi mu nzira y’ivyatsi vy’ibimera n’ingaruka zavyo ku mibiri yihariye The overall is. amakuru yerekeye umuntu ku giti ciwe. Iryo sesangura ryemeje kandi ko uruja n’uruza rw’imitsi y’imbere mu mubiri n’izindi mitsi y’ibimera atari JAs ari rwo ruhingura uburyo bwihariye bwo gusubira gutunganya ivy’umubiri bitewe n’ibikoko birya ivyatsi. Cane cane, twabonye ko ET yatewe na OSM yari nini cane kuruta iyo yatewe na OSS1. Ubwo buryo burahuye n’ibintu vyinshi birimwo FAC muri OSM, ivyo bikaba ari ibintu bikenewe kandi bihagije kugira ngo umuntu ashobore gutuma ET iturika (48). Mu bijanye n’ugukorana hagati y’ibimera n’ibikoko birya ivyatsi, igikorwa co gutanga ikimenyetso ca ET ku bijanye n’inguvu z’ibinyabuzima bitandukanye kiracari gitoyi kandi kigamije gusa umugwi umwe w’ibinyabuzima. Ikindi, inyigisho nyinshi zakoresheje gukoresha exogenous ET canke ibintu biyibanza canke inhibiteurs zitandukanye kugira ngo zige ingene ET itunganywa, muri ivyo ivyo gukoresha exogenous imiti bizoteza ingaruka nyinshi zidasanzwe. Ku bumenyi bwacu, iki cigwa kigereranya isuzuma rya mbere ry’urugero runini ry’uruhara rwa ET mu gukoresha ET mu guhingura no gutahura ibiterwa bihinduwe mu buryo butari bwo kugira ngo bihuze ubushobozi bw’ibimera. Ivyimburwa vy’ibikoko birya ivyatsi birashobora guhindura inyishu y’ibihimba vy’umubiri. Igihambaye cane ni ugukoresha ubuhinga bwa ET biosynthesis (ACO) n’ubuhinga bwo gutahura (ETR1) vyahishuye ivyimburwa vy’ibikoko birya ivyatsi vy’ibinyabuzima bita phenolamides. Vyerekanwe mbere ko ET ishobora gutunganya neza ukwirundanira kwa nikotine iterwa na JA mu gutunganya putrescine N-methyltransferase (49). Ariko rero, mu bijanye n’ubuhinga, ntibitomoye neza ingene ET itunganya neza ivy’ugutera umuti witwa phenamide. Uretse igikorwa co guhindura ikimenyetso ca ET, umuvuduko w’ibifungurwa urashobora kandi guhindurwa ukaja muri S-adenosyl-1-methionine kugira ngo utunganye ishoramari mu bihimba vy’umubiri bita polyaminophenol amides. S-adenosyl-1-methionine ni ET kandi ni ikintu gisanzwe kiri hagati y’inzira y’ubuhinga bw’ibinyabuzima. Uburyo ikimenyetso ca ET gitunganya urugero rwa phenolamide burakeneye kwigwa cane.
Kuva kera, kubera umubare munini w’ibinyabuzima bidasanzwe bifise imiterere itazwi, ukwitwararika cane ivyiciro vy’ibinyabuzima vyihariye ntikwashoboye gusuzuma neza amahinduka y’igihe y’ubudasa bw’ibinyabuzima inyuma y’ugukorana kw’ibinyabuzima. Muri iki gihe, bishingiye ku gusesangura ivyiyumviro vy’amakuru, igisubizo nyamukuru co kuronka urugero rwa MS/MS rushingiye ku bihinduka bitagira aho bigarukira ni uko ibikoko birya ivyatsi birya canke bigana ibikoko birya ivyatsi bibandanya kugabanya ubwoko bwose bw’ibifungurwa vy’ibinyabuzima vy’amababi mu gihe vyongera urugero rw’ubuhinga bwavyo. Ukwo kwongerekana kw’igihe gito kw’ubuhinga bwa metabolome buterwa n’ibikoko birya ivyatsi bifitaniye isano n’ukwiyongera kw’ubuhinga bwa transcriptome. Ikintu gifasha cane muri iyo metabolome nini (ifise agaciro ka Si kanini) ni metabolite idasanzwe ifise igikorwa co kurya ivyatsi cari caragaragajwe mbere. Ico kigereranyo gihuye n’ivyo OD ivuga, ariko ivyo MT ivuga bijanye n’ugusubira gukora porogarama y’ibihimba vy’umubiri ntibihuye. Ariko kandi, ayo makuru arahuye n’ivyo ubuhanuzi bw’ikigereranyo c’ibintu bivanze (MT nziza; Igishushanyo 1B), kubera ko ibindi bihimba vy’umubiri bitagira ibimenyetso bifise ibikorwa vy’ukwikingira bitazwi bishobora kuba bikiriko birakurikirana ugusangira kwa Si mu buryo butari bwo.
Icitegererezo gihambaye cafashwe n’ubu bushakashatsi ni uko kuva ku rugero rw’ubwihindurize buto (igiterwa kimwe n’itabi) gushika ku rugero runini rw’ubwihindurize (ubwoko bw’itabi bufitaniye isano rya hafi), ingero zitandukanye z’imitunganirize y’ubwihindurize ziri mu “kwikingira neza” Hariho itandukaniro rikomeye mu bushobozi bw’ibikoko birya ivyatsi. Moore n'abandi. (20) na Kessler na Kalske (1) bigenjeje neza ko bohindura ingero zitatu zikora z’ubudasa bw’ibinyabuzima zatandukanijwe mu ntango na Whittaker (50) zikaba amahinduka y’igihe y’ubudasa bw’ibinyabuzima; abo banditsi ntibavuze mu ncamake Uburyo bwo gukusanya amakuru y’ibinyabuzima ku rugero runini na bwo nyene ntibuvuga ingene umuntu yoharura ubudasa bw’ibinyabuzima bushingiye kuri ayo makuru. Muri iki cigwa, uguhindura gutoyi ku bijanye n’ugushikiriza ibikorwa kwa Whittaker kuzobona ubudasa bw’ibinyabuzima α nk’ubudasa bw’imirongo ya MS/MS mu giterwa kinaka, n’ubudasa bw’ibinyabuzima β nk’ubudasa bw’ibinyabuzima bw’ishimikiro bw’umugwi w’abantu.
Ico kimenyetso ca JA ni ngombwa ku nyishu nyinshi z’ibikoko birya ivyatsi. Ariko rero, nta n’igerageza rikomeye ry’ingero ry’uruhara rw’itegeko ry’ibinyabuzima vya JA ku bijanye n’ubudasa bw’ibinyabuzima, kandi nimba ikimenyetso ca JA ari ahantu rusangi h’ubudasa bw’ibinyabuzima buterwa n’imihangayiko ku rugero rwo hejuru rw’iterambere ry’ibinyabuzima biracari bigoye gutahura. Twabonye ko kamere y’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko vy’ibikoko. Ikindi, iyo ikimenyetso ca JA gihungabanye, ubuhinga bwo guhindura ibiribwa buterwa n’inyamaswa imwe y’umubiri birya ivyatsi buzokurwaho (Ishusho 3, C na E). Kubera ko amahinduka y’imiterere y’ibifungurwa vy’ibikoko vy’ibimera vy’ibimera vy’ibimera vy’ibimera vy’ibimera vy’ibimera vy’ibimera vy’ibimera vy’ibimera vy’ibimera vy’ibimera vy’ibimera vy’ibimera vy’ibinyabuzima vy’ibinyabuzima vy’ibinyabuzima vy’ibinyabuzima vy’ibinyabuzima vy’ibinyabuzima vy’ibinyabuzima. Ivyo bice vy’ibihimba vy’umubiri biganzwa n’igice c’umubiri kandi bikajana ku bijanye n’isano ryiza n’ibimenyetso vya JA.
Kubera ko uburyo bw’ibinyabuzima bw’ubwoko bw’itabi bufitaniye isano rya hafi na ryo butandukanye cane, ivyo bihimba bihinduka mu mubiri biramenyekana canecane mu bijanye n’uburyo, ni co gituma bishobora gusesangura cane. Ivyiyumviro vy’amakuru bikora ku bijanye n’imiterere y’ibifungurwa vyafashwe vyerekana ko ugutera ivyatsi bituma habaho uguhuza hagati y’ubudasa bw’ibifungurwa n’ubuhinga bwihariye. Ikimenyetso ca JA kirafise uruhara runini muri ivyo bihuza. Ukwongerekana kw’ubuhinga bwa metabolome guhuye n’ivyo OD ivuga kandi bifitaniye isano n’ikimenyetso ca JA, mu gihe ikimenyetso ca JA gifitaniye isano ribi n’ubudasa bw’imibiri. Ivyo bigereranyo vyerekana ko ubushobozi bwa OD bw’ibimera bugenwa ahanini n’ubuhinga bwa JA, haba ku rugero rw’iterambere rito canke ku rugero runini rw’iterambere. Igerageza ry’ugukoresha JA ry’inyuma ry’umubiri ryikingira utunenge tw’ibinyabuzima vya JA ryerekana kandi ko ubwoko bw’itabi bufitaniye isano rya hafi bushobora gutandukanywa mu bwoko bwishura ku kimenyetso n’ubwoko butasubiza ku kimenyetso, nk’uko nyene uburyo bwavyo bwo gukora JA n’ubuhinga bwa metabolome buterwa n’ibikoko birya ivyatsi. Ubwoko butagira inyishu y’ibimenyetso ntibushobora kwishura kubera ukudashobora kwavyo gutanga JA y’imbere mu mubiri kandi rero burashobora gushikirwa n’imipaka y’umubiri. Ivyo bishobora guterwa n’uguhinduka kw’ibihimba bimwebimwe vy’ingenzi mu nzira y’ibimenyetso vya JA (AOS na JAR4 muri N. crescens) y’. Ico ciyumviro kirerekana ko izo nzira z’iterambere ry’ibinyabuzima zishobora guterwa ahanini n’amahinduka mu kuntu umuntu abona ivy’imbere mu mubiri be n’ingene yishura.
Uretse ugukorana hagati y’ibimera n’ibikoko birya ivyatsi, ugutohoza ubudasa bw’ibifungurwa birafitaniye isano n’iterambere ryose rihambaye ry’ivyiyumviro mu bushakashatsi bw’ukuntu ibinyabuzima bimenyerera ibidukikije be n’ugutera imbere kw’ibiranga ibinyabuzima bikomeye. Kubera ko amakuru abonwa n’ibikoresho vya MS vya none yiyongera, ubu igerageza ry’ivyiyumviro ku bijanye n’ubudasa bw’ibifungurwa birashobora kurenga itandukaniro ry’ibifungurwa vy’umuntu ku giti ciwe/ivy’imigwi maze rigakora isesengura ry’isi yose kugira ngo rigaragaze uburyo butategerezwa. Mu nzira y’isesengura ry’urugero runini, imvugo ngereranyo ihambaye ni iciyumviro co gufata amakarata afise insiguro ashobora gukoreshwa mu gutohoza amakuru. Rero, igisubizo gihambaye c’uguhuza ubu ubuhinga bw’imibiri n’inyigisho y’amakuru bitagira aho bigarukira ni uko bitanga ubuhinga bworoshe bushobora gukoreshwa mu kwubaka amakarata kugira ngo umuntu asuzume ubudasa bw’imibiri ku rugero rutandukanye rw’imibiri. Ni co kintu nyamukuru gikenewe muri ubwo buryo. Ivyigwa vy’iterambere ry’ibintu bitobito/bikomeye n’ibidukikije vyo mu kibano.
Ku rugero rw’iterambere rikomeye, ishingiro ry’inyigisho y’iterambere ry’ibimera n’udukoko rya Ehrlich na Raven (51) ni ukuvuga imbere y’igihe ko uguhinduka kw’ubudasa bw’imibiri hagati y’ubwoko ari bwo butera uguhinduka kw’imiryango y’ibimera. Ariko rero, mu myaka mirongo itanu kuva iki gitabu c’imbuto gisohotse, iyo nkuru ntiyageragejwe cane (52). Ivyo ahanini biva ku biranga phylogenetic vy’ibiranga ibiribwa bigereranywa mu miryango y’ibimera biva kure cane. Ivyo bishobora gukoreshwa mu gushingira intahe uburyo bwo gusesangura intumbero. Ivyo MS/MS ikora ubu bikoreshwa n’inyigisho y’amakuru bipima ukuntu MS/MS isa n’ibindi bihimba bitazwi (ata guhitamwo imbere y’igihe) maze bikahindura ivyo bihimba vy’umubiri bikaba umugwi wa MS/MS, gutyo mu guhindura ibihimba vy’umubiri vy’umwuga Ivyo bigereranyo vy’iterambere rikomeye biragereranywa mu rugero rwo gushikiriza. Ivyerekana imibare vyoroshe. Ivyo bikorwa bisa n’isesengura ry’ivy’ubuhinga bwa filogenetike, rishobora gukoresha urutonde rw’ibintu kugira ngo rimenye urugero rw’uguhinduka canke uguhinduka kw’ibiranga ataco bivuze imbere y’igihe.
Ku rugero rw’ubuhinga bw’ibinyabuzima, iciyumviro co gusuzuma ca Firn na Jones (53) kirerekana ko ubudasa bw’ibinyabuzima buguma ku rwego rutandukanye kugira ngo haboneke ibikoresho vya mbere vyo gukoresha ibikorwa vy’ibinyabuzima vy’ibinyabuzima vyari bitagira isano canke vyasubirijwe. Uburyo bw’inyigisho y’amakuru buratanga urutonde aho izo mpinduka zijanye n’iterambere ry’ibihimba vy’umubiri zibaho mu gihe c’ubuhinga bw’ibihimba vy’umubiri zishobora guharurwa nk’igice c’urugendo rwo gusuzuma iterambere ry’ibihimba vy’umubiri: uguhinduka kw’ibinyabuzima kuva ku buhinga buke gushika ku bwihariye bwinshi Ibihimba vy’umubiri bibujijwe vy’ibidukikije bimwebimwe.
Muri vyose, mu misi ya mbere y’ubuhinga bw’ibinyabuzima, harashizweho inyigisho zihambaye zo kwikingira ibimera, kandi uburyo bushingiye ku vyiyumviro bishingiye ku vyo umuntu abona burabonwa cane ko ari bwo buryo bwonyene bwo gutera imbere mu vya siyansi. Ivyo ahanini biterwa n’ubuhinga bwo gupima metabolome yose. Naho uburyo bushingiye ku vyiyumviro ari ngirakamaro cane mu guhitamwo ubundi buryo butera, ubushobozi bwabwo bwo guteza imbere ugutahura kwacu ku bijanye n’imirongo y’ibinyabuzima burake cane kuruta uburyo bwo gukoresha ubuhinga bw’ibarabara buriho ubu mu buhinga bwo muri iki gihe bukoresha amakuru menshi. Rero, ivyiyumviro bidashobora kumenyekana birarengeye kure cane urugero rw’amakuru ariho, rero umurongo w’ivyiyumviro/ingendo y’iterambere mu bijanye n’ubushakashatsi ntushobora gukurwaho (4). Turabona ko ubuhinga bwo gukoresha ubuhinga bwa metabolomics bwashizwe aha bushobora gusubira gutuma abantu bakunda ibibazo biherutse (ingene) n’ivya nyuma (kubera iki) vy’ubudasa bw’ibifungurwa, kandi bikagira ico bikoze ku gihe gishasha c’ubuhinga bw’amakuru burongowe n’ivyiyumviro. Ico gihe carasubiyemwo inyigisho zihambaye zatumye amayaruka ya kera agira ico akora.
Ugufungura ibikoko birya ivyatsi bitaziguye bikorwa mu kurera igiterwa ca kabiri c’instar canke igiterwa ca Sl ku ibabi rimwe ry’igitegwa c’ibara ry’agahama ry’igiterwa kimwe gitera amashurwe y’urugori, hakaba hariho ibiterwa 10 vyisubiriza ku giterwa. Ivyo bikoko vyafatwa n’ibifatanya, maze amababi yasigaye akakwegeranywa amasaha 24 n’amasaha 72 inyuma y’aho arwaye maze agakonja ningoga, hanyuma ivyo bihimba vy’umubiri bikakurwamwo.
Kwigana ubuvuzi bw’ibikoko birya ivyatsi mu buryo buhuye cane. Uburyo ni ugukoresha amakurudumu y’impuzu kugira ngo utobore imirongo itatu y’amahwa ku ruhande rumwe rumwe rw’urubavu rwo hagati rw’amababi atatu y’igitegwa yagutse neza mu gihe c’ukukura kw’igitsibo c’impuzu, hanyuma ugaca ukoresha 1:5 Diluted Ms. Canke ukoreshe intoke zifise amaganzu kugira ngo utobore S1 OS mu gipfukisho. Kwimbura no gukora ibabi nk’uko vyavuzwe haruguru. Koresha uburyo bwavuzwe imbere kugira ngo ukuremwo ibintu vy’intango bihindura ibiribwa n’imitsi y’ibimera (54).
Ku bikorwa vya JA biva hanze, amababi atatu y’amababi ya buri kimwe mu bimera bitandatu vy’amashurwe y’amaroza vy’ubwoko bumwe bumwe avuzwa n’imiti 20 μl y’umunyu wa lanoline urimwo 150μg ya MeJA (Lan + MeJA), na 20μl ya lanoline hamwe n’umuti wo gucungera ibikomere (Lan + la2), canke ukoresheje pur. Amababi yasarurwa amasaha 72 inyuma y’aho avuwe, agakonja mu mazi y’azote, hanyuma akabikwa ku bushuhe bwa -80°C gushika akoreshejwe.
Imirongo ine y’ibihimba vy’umubiri bihinduka, ari yo irAOC (36), irCOI1 (55), irACO na sETR1 (48), yarabonetse mu mugwi wacu w’ubushakashatsi. irAOC yerekanye cane ko JA na JA-Ile zigabanutse, mu gihe irCOI1 itagira ico ikora ku JAs. Ugereranije na EV, ukwirundanira kwa JA-Ile kwariyongereye. Na irACO nyene izogabanya ivyimburwa vya ET, kandi ugereranije na EV, sETR1, idashobora gutahura ET, izokwongerera ivyimburwa vya ET.
Igikoresho co gupima umuco w’izuba (Sensor Sense ETD-300) gikoreshwa mu gupima ET ataco umuntu akora. Ubwo nyene inyuma y’aho amababi avuwe, igice c’amababi caratemwa, kigashirwa mu gitereko c’ibirahuri c’imiliyoni 4 gipfutse, maze umwanya wo mu mutwe ukarekurwa ngo wirundanire mu masaha 5. Mu gihe c’ingero, igikombe cose carasukuwe n’umugezi w’umuyaga utagira agasembwa w’amalita 2/isaha mu minota 8, uwo muyaga wari waraciye mbere mu gikoresho co gutuma CO2 n’amazi bikuramwo.
Amakuru ya microarray yasohowe ubwa mbere mu (35) maze abikwa mu kigo c’igihugu c’amakuru y’ubuhinga bwa none (NCBI) c’amakuru y’ivy’imiterere y’imitsi (inomero y’uruja n’uruza GSE30287). Amakuru ahuye n’amababi yatewe n’ubuvuzi bwa W + OSMs n’ubugenzuzi butagiramwo ivyago vyakuwe muri iki cigwa. Inkomezi zidasanzwe ni log2. Imbere y’isesengura ry’imibare, umurongo w’intango warahinduwe maze urasanzwe ku rugero rwawo rwa 75 kw’ijana hakoreshejwe porogarama ya R.
Amakuru y’intango y’urutonde rwa ARN (RNA-seq) y’ubwoko bwa Nicotiana yaronswe mu bubiko bw’ibisomwa bigufi bwa NCBI (SRA), inomero y’umugambi ni PRJNA301787, yatangajwe na Zhou n’abandi. (39) hanyuma ugende nk’uko bivugwa muri (56). Amakuru atagiramwo ivyatsi yatunganijwe na W + W, W + OSM na W + OSS1 ahuye n’ubwoko bwa Nicotiana yatowe kugira ngo asesengurwe muri iri koraniro, kandi atunganijwe mu buryo bukurikira: Ubwa mbere, ibisomwa bitagiramwo ivyatsi vya RNA-seq vyahinduwe mu buryo bwa FASTQ. HISAT2 ihindura FASTQ mu SAM, na SAMtools ihindura amadosiye ya SAM mu madosiye ya BAM yatoranijwe. StringTie ikoreshwa mu kubara insobanuro y’uruvyaro, kandi uburyo bwayo bwo kugaragaza ni uko hariho ibice ku bice igihumbi vy’ishimikiro ku miliyoni y’ibice vy’inyandiko bikurikiranye.
Inkingi ya Acclaim (150 mm x 2,1 mm; ubunini bw’ibice 2,2μm) yakoreshejwe mu gusesangura n’inkingi y’uburinzi ya 4 mm x 4 mm bikozwe mu kintu kimwe. Ivyo bimenyetso bibiri bikurikira bikoreshwa mu buhinga bwo gupima amazi bufise ubushobozi bwinshi cane (UHPLC): 0 gushika ku minota 0,5, isocratic 90% A [amazi atagira iyoni, 0,1% (v/v) acetonitrile (v/v) acetonitrile na 0,05% B 0,05% vy’aside formique); 0,5 gushika ku minota 23,5, igice c’ibara ry’umubiri ni 10% A na 90% B, uko bikurikirana; 23,5 gushika ku minota 25, isocratique 10% A na 90% B. Igitigiri c’amazi atemba ni 400μl/min. Ku bijanye n’isesengura ryose rya MS, ushire amazi y’inkingi mu gikoresho co gusesangura c’amabara ane n’igihe co guguruka (qTOF) gifise isoko y’umuyagankuba ikora mu buryo bw’iyonizasiyo nziza (umuriro w’imitsi, 4500 V; isohoka ry’imitsi 130 V ; Ubushuhe bwo kwuma 200°C liters ai/10).
Gukora isesengura ry’ibice vya MS / MS (ubu ryitwa MS / MS) ritagira akamaro canke ridashobora gutandukanywa n’amakuru kugira ngo uronke amakuru y’imiterere yerekeye imiterere y’imibiri muri rusangi ishobora kumenyekana. Iciyumviro c’uburyo bwa MS/MS butagira ivangura gishingiye ku kuba iyo quadrupole ifise idirisha rinini cane ry’ukwitandukanya n’uburemere [ni co gituma, uzirikane ibimenyetso vyose vy’uburemere n’uburemere (m/z) nk’ibice]. Kubera iyo mpamvu, kubera ko igikoresho ca Impact II kitashoboye kurema uguhengamira kwa CE, hakozwe isesengura ryinshi ryigenga hakoreshejwe agaciro k’inguvu z’ugusenyuka kw’ugusenyuka (CE) ziterwa n’ugutombora. Muri make, banza usesengure igipimo hakoreshejwe UHPLC-electrospray ionization/qTOF-MS ukoresheje uburyo bumwe bwo gupima uburemere bw’ibintu (ibintu bike vy’uguca ibice biva ku guca ibice biri mu nzira), ukoresheje gucapura kuva kuri m/z 50 gushika kuri 1500 ku nkuru y’ugusubiramwo 5 Hz. Koresha azote nk’umuyagankuba w’ugutombora mu gusesangura MS/MS, kandi ukore ingero zigenga kuri izo nzira zine zitandukanye z’ugusenyuka ziterwa n’ugutombora: 20, 30, 40, na 50 eV. Mu gihe cose c’ugupima, iyo quadrupole irafise idirisha rinini ry’ugutandukanya uburemere, kuva kuri m/z 50 gushika kuri 1500. Iyo umubiri w’imbere m/z n’igerageza ry’ubwaguke bw’ugutandukanya bishizwe kuri 200, urutonde rw’uburemere ruca rukora ubwarwo n’ubuhinga bwo gukoresha igikoresho na 0 Da. Scan ku bice vy’uburemere nk’uko biri mu buryo bw’uburemere bumwe. Koresha umuti w’amazi (50 ml ya isopropanol, 200 μl ya aside formique na militiro 1 y’umunyu w’amazi wa NaOH 1M) kugira ngo ugereranije uburemere bw’ibintu. Hakoreshejwe ubuhinga bwo gupima bufise ubuhinga bwinshi bwa Bruker, dosiye y’amakuru irapimwa inyuma yo gukoresha urugero rwa mwayeni mu kiringo kinaka. Koresha igikorwa co kohereza hanze porogarama y’Isesengura ry’Amakuru v4.0 (Brook Dalton, Bremen, Ubudage) kugira ngo uhindure amadosiye y’amakuru atagiramwo ivyatsi mu buryo bwa NetCDF. Ivyerekeye amakuru ya MS/MS vyabitswe mu rutonde rw’amakuru rw’ivy’imibiri rwuguruye rwitwa MetaboLights (www.ebi.ac.uk) rufise inomero y’ugushiramwo. MTB1471.
Iteraniro rya MS/MS rishobora gushikako biciye mu gusesangura isano hagati y’ibimenyetso vy’uburyo bwa MS1 na MS/MS ku nguvu z’ugutombora ziri hasi n’iziri hejuru be n’amategeko mashasha yashizwe mu ngiro. Inyandiko ya R ikoreshwa mu gushitsa isesengura ry’isano ry’ugusangira kw’imbere y’igicuruzwa, kandi inyandiko ya C# (https://github.com/MPI-DL/indiscriminant-MS-MS-assembly-pipeline) ikoreshwa mu gushitsa amategeko.
Kugira ngo tugabanye amakosa meza y’ibinyoma aterwa n’urusaku rw’inyuma n’uguhuza kw’ibinyoma guterwa no kumenya ibiranga m/z bimwebimwe mu bigereranyo bikeyi gusa, dukoresha igikorwa ca “filled peak” c’umugwi wa R XCMS (ku bijanye no gukosora urusaku rw’inyuma) Ikwiye gukoreshwa mu gusubirira “petende peak” (petende peak”). Iyo igikorwa co kwuzuza isonga gikoreshwa, haracariho agaciro kenshi “0′′ k’uburemere mu rutonde rw’amakuru ruzogira ico rukoze ku kubara kw’ihuriro. Hanyuma, tugereranya ibisubizo vyo gutunganya amakuru vyabonetse igihe igikorwa c’isonga ryuzuye gikoreshwa n’igihe igikorwa c’isonga ryuzuye kitakoreshwa, maze tukabara agaciro k’urusaku rw’inyuma akosowe hakurikijwe ivyo agaciro k’uburemere n’agaciro k’inyuma kaharuwe Twarihweje kandi gusa ibiranga uburemere bwavyo burenze incuro zitatu agaciro k’inyuma maze tukabibona ko ari “isonga ry’ukuri.” Ku biharuro vya PCC, ibimenyetso vya m/z vy’imbere y’akarorero (MS1) n’ibice vy’amakuru bifise nibura isonga umunani z’ukuri ni vyo vyonyene vyiyumviriwe.
Iyo uburemere bw’ikintu c’uburemere bw’imbere mu citegererezo cose bufitaniye isano rikomeye n’uburemere bugabanutse bw’ico kintu nyene c’uburyo gishikirwa n’inguvu z’ugutomboka ziri hasi canke nyinshi, kandi ico kintu kitashizweko ikimenyetso c’uko ari isonga ry’isotope na CAMERA, birashobora gusobanurwamwo neza. Hanyuma, mu kubara ibice vyose bishoboka vy’imbere n’ibintu bibiri mu masegonda 3 (idirisha ry’igihe co kugumya ry’isonga), isesengura ry’isano rikorwa. Iyo agaciro ka m/z kari hasi y’agaciro k’imbere kandi uguca ibice kwa MS/MS kuba mu kibanza kimwe c’akarorero mu rutonde rw’amakuru n’imbere yakomotseko, ni ho gusa bifatwa nk’igice.
Hashingiwe kuri ayo mategeko abiri yoroshe, turakuraho ibice vyerekanwa bifise agaciro ka m/z karuta m/z k’imbere y’imbere yamenyekanye, kandi dushingiye ku kibanza c’akarorero aho imbere y’imbere igaragara n’igice cavuzwe. Birashoboka kandi guhitamwo ibiranga uburyo bivuye ku bice vyinshi vyo mu nkomoko bivuye mu buryo bwa MS1 nk’ibibanza vy’abahiganwa, gutyo bikazana ibirungo vya MS/MS bitari vyinshi. Kugira ngo ivyo bimenyetso bigabanuke, iyo ugusa kw’imirongo y’ibara ry’umuhondo (NDP) kurenga 0,6, kandi bikaba biri mu “pcgroup” y’ibara ry’umuhondo (chromatogram) ryanditswe na CAMERA, tuzobifatanya. Ubwa nyuma, dufatanya ibisubizo vyose bine vya CE bifitaniye isano n’imbere n’ibice mu rwego rwa nyuma rw’ibintu bihurikiye hamwe mu guhitamwo isonga ry’uburemere buhambaye kuruta ayandi yose mu isonga ry’abahiganwa rifise agaciro ka m/z kamwe ku nguvu zitandukanye z’ugutombora. Intambwe zikurikira zo gutunganya zishingiye ku ciyumviro c’ibara ry’ibintu vyinshi kandi zizirikana ibintu bitandukanye vya CE bisabwa kugira ngo umuntu ashobore guca ibice, kuko ibice bimwebimwe bishobora gusa kumenyekana munsi y’inguvu zimwe zimwe z’ugutombora.
RDPI (30) yakoreshejwe mu kubara ubushobozi bwo guhindura ibiribwa. Ubutandukane bw’imirongo y’ibinyabuzima (index ya Hj) buva ku bwinshi bw’ibibanza vya MS/MS hakoreshejwe entropie ya Shannon y’ugusangira kw’incuro za MS/MS hakoreshejwe iyi nsiguro ikurikira yadondowe na Martínez n’abandi. (8). Hj = −∑i = 1mPijlog2(Pij) aho Pij ihuye n’incuro zihuye n’ivyo MS/MS ya i mu citegererezo ca j (j = 1, 2,..., m) (i = 1, 2, ..., m) t).
Ubudasa bw’imibiri (Si index) busobanurwa nk’akaranga k’imvugo y’i MS/MS itangwa mu bijanye n’incuro hagati y’ibigereranyo biriko birasuzumwa. MS/MS ubudasa buharurwa nk'uko Si = 1t (∑j = 1tPijPilog2PijPi)
Koresha iyi formile kugira ngo upime urutonde rwa δj rwerekeye metabolome rw’akarorero ka j kose, n’umubare w’ibintu vy’agaciro vya MS/MS δj = ∑i = 1mPijSi .
Ivyiyumviro vya MS/MS bihuzwa bibiri bibiri, kandi ukuntu bisa biharurwa bishingiye ku manota abiri. Ubwa mbere, ukoresheje NDP isanzwe (izwi kandi nk’uburyo bwo guhuza cosine), ukoreshe iyi nsiguro ikurikira kugira ngo ushire amanota ku bijanye n’ugusa kw’igice hagati y’imirongo NDP = (∑iS1 & S2WS1, iWS2, i) 2∑iWS1, i2∑iWS2, i2 aho S1 na S2 nk’iriba Bihuye, ku mirongo nk’iWS2, 1. i igereranya uburemere bushingiye ku bukomezi bw’isonga ko itandukaniro ry’isonga rusangi rya i hagati y’ivyo bimenyetso bibiri riri munsi ya 0,01 Da. Uburemere buharurwa gutya: W = [uburemere bw’isonga] m [uburyo] n, m = 0,5, n = 2, nk’uko vyasabwe na MassBank.
Haciye hashirwa mu ngiro uburyo bwa kabiri bwo gutanga amanota, bwari bujanye no gusuzuma NL rusangi hagati ya MS/MS. Kugira ngo ivyo bishoboke, twakoresheje urutonde rwa NL 52 rwakunda guhura mu gihe c’uguca ibice kwa MS mu buryo bumwe, n’urutonde rwa NL rwihariye cane (dosiye y’amakuru S1) rwari rwarasobanuwe mbere ku bijanye n’urutonde rwa MS/MS rw’ibinyabuzima bibiri vy’ubwoko bwa Nepenthes bwagoyagoye ( 9, 26). Rema vecteur binaire ya 1 na 0 kuri buri MS/MS, ihuye n’ubu n’ibitabaho vya NL zimwe zimwe. Hashingiwe ku gusa kw’urugendo rwa Euclide, igiharuro c’ugusa kwa NL giharurwa ku bijanye n’imirongo ibiri ya NL.
Kugira ngo dukore amatsinda abiri, twakoresheje umugwi wa R DiffCoEx, ushingiye ku kwagura kw’isesengura ry’imiterere y’imitsi y’umubiri (WGCNA). Dukoresheje amabara y’ibiharuro vya NDP na NL vy’imirongo ya MS/MS, twakoresheje DiffCoEx kugira ngo tubaze amabara y’ubufatanye bw’ibigereranyo. Ivyiyumviro bibiri bikoreshwa mu gushiramwo “cutreeDynamic” parameter ku buryo = “ivy’ivyaduka”, cutHeight = 0.9999, deepSplit = T, na minClusterSize = 10. Kode y’inkomoko R ya DiffCoEx yavanwe muri dosiye y’inyongera 1 na Tesson n’abandi. (57); Ivyo bisabwa vy’ubuhinga bwa none vya R WGCNA biraboneka kuri https://horvath.genetics.ucla.edu/html.
Kugira ngo dukore isesengura ry’urusobe rw’amamolekule rwa MS/MS, twaharuriye ubufatanye bw’imirongo ibiri bushingiye ku bwoko bw’ugusa kwa NDP na NL, maze dukoresha porogarama ya Cytoscape kugira ngo tubone uko urusobe ruteye dukoresheje imiterere y’ibinyabuzima mu gikorwa co kwagura imiterere y’imirongo ya CyFilescape yFiles.
Koresha R verisiyo 3.0.1 kugira ngo ukore isesengura ry’imibare ku makuru. Ivyerekeye imibare vyasuzumwe hakoreshejwe uburyo bubiri bwo gusesangura ubudasa (ANOVA), hakurikijwe ikigeragezo ca Tukey c’inyuma y’igihe c’itandukaniro ry’agaciro (HSD). Kugira ngo hasuzumwe itandukaniro hagati y’ubuvuzi bw’ibikoko birya ivyatsi n’ubuvuzi bwo kugenzura, ugusangira kw’imirongo ibiri kw’imigwi ibiri y’ibipimo bifise itandukaniro rimwe kwasuzumwe hakoreshejwe ikigeragezo ca Student’s t.
Ku bijanye n’ibikoresho vy’inyongera vy’iyi ngingo, raba kuri http://advances.sciencemag.org/cgi/ibirimwo/vyuzuye/24/6/eaaz0381/DC1.
Iyi ni ingingo ishobora gukoreshwa n’abantu bose igakwiragizwa hakurikijwe amategeko y’uruhusha rwo gukoresha, gukwiragiza no gusubiramwo mu buryo ubwo ari bwo bwose, igihe cose ikoreshwa rya nyuma ritari ryo gushaka inyungu z’ubudandaji kandi ikintu nyamukuru ari uko igikorwa c’intango ari co. Urutonde.
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