Waad ku mahadsan tahay booqashada Nature.com. Nooca biraawsarka aad isticmaaleyso wuxuu leeyahay taageero CSS oo xaddidan. Waayo-aragnimada ugu fiican, waxaan kugula talineynaa inaad isticmaasho biraawsar la cusbooneysiiyay (ama aad joojiso Habka La-qabsiga ee Internet Explorer). Dhanka kale, si loo hubiyo taageerada joogtada ah, waxaan soo bandhigi doonnaa goobta iyada oo aan lahayn qaabab iyo JavaScript.
Angustifolius lupine (NLL, Lupinus angustifolius L.) waa geed leguminous ah oo loo isticmaalo wax soo saarka cuntada iyo horumarinta carrada. Ballaarinta caalamiga ah ee NLL sida dalagga ayaa soo jiidatay fangaska badan ee cudurada keena, oo ay ku jiraan lupine anthracnose, kaas oo sababa cudurka anthracnose ee ba'an. Laba alleles, Lanr1 iyo AnMan, kuwaas oo bixiya iska caabin kordhay, ayaa loo isticmaalay taranka NLL, laakiin farsamooyinka molecular-ka ee hoose weli lama oga. Daraasaddan, calaamadaha Lanr1 iyo AnMan ayaa loo isticmaalay in lagu baaro muunadaha NLL ee Yurub. Tijaabada tallaalka ee jawi la xakameeyey ayaa xaqiijisay waxtarka labada deeq-bixiye ee iska caabiya. Tilmaanta kala duwan ee muujinta hidda-wadaha ayaa lagu sameeyay xariiqyo u adkaysta wakiillada iyo kuwa u nugul. Iska caabbinta Anthracnose waxay la xiriirtay muujinta xad-dhaafka ah ee ereyada ontology-ga hiddaha "GO:0006952 Jawaabta Difaaca", "GO:0055114 Habka Redox", iyo "GO:0015979 Sawir-qaadista". Intaa waxaa dheer, khadka Lanr1(83A:476) wuxuu muujiyay dib-u-habayn muhiim ah oo ku saabsan qoraalka ka dib tallaalka, halka khadadka kale ay muujiyeen dib u dhac ku yimid jawaabtan qiyaastii 42 saacadood. Jawaabaha difaaca waxay la xiriiraan hiddo-wadaha TIR-NBS, CC-NBS-LRR iyo NBS-LRR, 10 borotiin oo ku lug leh cudurka, borotiinnada wareejinta lipid, endoglucan-1,3-β-glucosidase, borotiinnada derbiga unugyada hodanka ku ah glycine, iyo hiddo-wadaha ka yimaada wadada falcelinta ee oksijiinta. Jawaabaha hore ee 83A:476, oo ay ku jiraan xakamaynta taxaddar leh ee hiddo-wadaha la xiriira sawir-qaadista, waxay la jaanqaadeen ilaalin guul leh inta lagu jiro marxaladda koritaanka dhirta ee bayoolajiga fungal, taasoo soo jeedinaysa in saameyn-bixiye uu kiciyo difaac. Falcelinta Mandeloop waa la gaabiyaa, sidoo kale jiidista guud ee toosan.
Lupine-ka caleenta cidhiidhiga ah (NLL, Lupinus angustifolius L.) waa badar borotiin badan leh oo ka soo jeeda gobolka galbeedka Mediterranean1,2. Hadda waxaa loo beeraa sidii dalag cunto ah oo loogu talagalay xayawaanka iyo aadanaha. Waxaa sidoo kale loo arkaa digada cagaaran ee nidaamyada wareegga dalagga sababtoo ah hagaajinta nitrogen-ka iyadoo la adeegsanayo bakteeriyada isku-dhafan ee hagaajisa nitrogen-ka iyo horumarinta guud ee qaab-dhismeedka ciidda. NLL waxay soo martay geeddi-socod degdeg ah oo ku saabsan daaqsinka qarnigii la soo dhaafay waxayna weli ku jirtaa cadaadis taran oo sarreeya3,4,5,6,7,8,9,10,11,12. Iyadoo la beerayo NLL ballaaran, isku-darka fangaska cudurada keena waxay sameeyeen beero cusub waxayna sababeen cudurro cusub oo burburiya dalagga. Waxa ugu yaabka badan beeralayda iyo kuwa wax tarta lupin-ka waxay ahayd muuqaalka anthracnose, oo uu sababay fangaska faafa, Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Waxa ugu yaabka badan beeralayda iyo kuwa wax tarta lupin-ka waxay ahayd muuqaalka anthracnose, oo uu sababay fangaska faafa, Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Наиболее примечательным для фермеров и селекционеров. Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Waxa ugu caansan beeralayda iyo kuwa wax tarbiya lupine-ka waxay ahayd soo ifbixidda anthracnose oo uu sababay fangaska bakteeriyada keena ee Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13.对于羽扇豆农民和饲养者来说，最引人注目的是炭疽病的出现，它是由病原真語是由病原真真真真看Feedi (Botricle) Hagedorn13 引起的.对于羽扇豆农民和饲养者来说，最引人注目的是炭疽病的出现。 Наиболее поразительным для фермеров грибком Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Waxa ugu yaabka badan beeralayda iyo kuwa wax tarta lupine waa soo ifbaxa anthracnose oo uu keeno fangaska bakteeriyada keena ee Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13.Warbixinnadii ugu horreeyay ee cudurkan waxay ka yimaadeen Brazil iyo Mareykanka, iyadoo calaamadaha caadiga ah ay soo baxeen 1912 iyo 1929 siday u kala horreeyaan. Si kastaba ha ahaatee, ka dib qiyaastii 30 sano, cudurka waxaa loo magacaabay Colletotrichum gloeosporioides (Penz.) Penz. & Sacc., teleomorph Glomerella cingulata (Stoneman) Spauld. & Sacc., teleomorph Glomerella cingulata (Stoneman) Spauld. & Sacc., телеоморф Glomerella cingulata (Stoneman) Spauld. & Sacc., teleomorph ee Glomerella cingulata (Stoneman) Spauld. & Sacc.，有目的形态的 Glomerella cingulata (Stoneman) Spauld. & Sacc.，有目的形态的 Glomerella cingulata (Stoneman) Spauld. & Sacc., Glomerella cingulata (Stoneman) Spauld в Целенаправленной морфологии. & Sacc., Glomerella cingulata (Stoneman) Spauld oo ku jira qaab-dhismeedka bartilmaameedka. & H. Schrenk,. & H. Schrenk,.iyo H. Schrenk. & H.施伦克，. & H.施伦克，.iyo H. Schlenk,.Tijaabinta cudurka ee hordhaca ah ee la sameeyay bartamihii qarnigii 20-aad waxay muujisay iska caabin NLL iyo lupine huruud ah (L. luteus L.), laakiin dhammaan isku-darka lupine cad (L. albus L.) ee la tijaabiyay waxay ahaayeen kuwo aad u nugul15,16. Daraasaduhu waxay muujiyeen in horumarinta anthracnose ay la xiriirto roobab badan (qoyaanka hawada) iyo heerkulka (inta u dhaxaysa 12-28°C), taasoo keentay jebinta iska caabinta heerkulka sare17, 18. Xaqiiqdii, waqtiga loo baahan yahay in conidia ay biqsho oo cudurku bilaabmo, wuxuu ahaa afar jeer ka gaaban 24°C (4 saacadood) marka loo eego 12°C (16 saacadood) xaaladaha qoyaanka sare19. Sidaa darteed, kulaylka caalamiga ah ee socda ayaa horseeday faafitaanka anthracnose. Si kastaba ha ahaatee, cudurka waxaa lagu arkay Faransiiska (1982) iyo Yukrayn (1983) isagoo ah calaamad muujinaysa khatar soo socota, laakiin sida muuqata waxaa iska indhatiray warshadaha lupine waqtigaas20,21. Dhawr sano ka dib, cudurkan ba'an wuxuu ku faafay adduunka oo dhan wuxuuna sidoo kale saameeyay waddamada waaweyn ee soo saara lupine sida Australia, Poland iyo Germany22,23,24. Ka dib markii uu faafay cudurka anthracnose bartamihii 1990-meeyadii, baaritaan ballaaran ayaa sababay in la aqoonsado dhowr deeq-bixiye oo iska caabiya muunadaha NLL19. Iska caabbinta NLL ee anthracnose waxaa xakameeya laba alleles oo kala duwan oo laga helo ilo kala duwan oo jeermiska ah: Lanr1 oo ku jira noocyada kala duwan ee Tanjil iyo Wonga iyo AnMan oo ku jira noocyada kala duwan. Mandalay 25, 26. Alleles-kani waxay dhammaystiraan calaamadaha molecular-ka ee taageera xulashada jeermiska u adkaysta barnaamijyada taranka25, 26, 27, 28, 29, 30. Khadka taranka ee u adkaysta 83A:476 oo sita allele-ka Lanr1 ayaa laga gudbay khadka duurjoogta ah ee u nugul P27255 si loo helo tirada RIL oo kala soocda iska caabbinta anthracnose, taasoo suurtogal ka dhigtay in loo qoondeeyo goobta Lanr1 koromosoomka NLL-1131, 32, 33. Isku-dubaridka calaamadaha khariidadda isku xidhka laga bilaabo meesha iska caabbinta ee dhinacyada ilaa anthracnose iyadoo la adeegsanayo qaab-dhismeedka hidde-sideyaasha, NLL waxay shaaca ka qaadday goobta dhammaan saddexda alleles ee ku yaal isla koromosoomyada (NLL-11), laakiin boosas kala duwan29,34,35. Si kastaba ha ahaatee, sababtoo ah tirada yar ee RILs iyo masaafada hidde-sideyaasha ee weyn ee u dhaxaysa calaamadaha iyo alleles-ka u dhigma, ma jiraan wax gunaanad ah oo lagu kalsoonaan karo oo ku saabsan hidde-sideyaashooda hoose. Dhanka kale, isticmaalka hidde-sideyaasha dib-u-soo-noqoshada ee lupins-ku waa adag tahay sababtoo ah awooddooda dib-u-soo-noolaynta oo aad u hooseysa, taasoo ka dhigaysa maaraynta hidde-sideyaasha mid adag37.
Horumarinta jeermiska la beeray ee sida allele-ka la rabo ee xaaladda homozygous, sida 83A:476 (Lanr1) iyo Mandelup (AnMan), ayaa albaabka u furtay barashada iska caabbinta anthracnose iyadoo la eegayo jiritaanka isku-darka iska soo horjeeda ee alleles ee dadka duurjoogta ah. Suurtagalnimada hababka molecular-ka. Isbarbardhig jawaabaha difaaca ee ay soo saaraan genotypes gaar ah. Daraasaddani waxay qiimaysay jawaab celinta hore ee qoraalka ee NLL ee tallaalka C. lupini. Marka hore, guddi germplasm ah oo Yurub ah oo NLL ah oo ka kooban 215 xariiq ayaa la baaray iyadoo la adeegsanayo calaamado molecular ah oo calaamadeeya alleles-ka Lanr1 iyo AnMan. Dabadeed waxaa lagu sameeyay 50 xariiq oo NLL ah, oo hore loogu xushay calaamadaha molecular-ka, xaaladaha la xakameeyey. Iyada oo lagu saleynayo tijaabooyinkan, afar xariiq oo kala duwan oo ku kala duwan iska caabbinta anthracnose iyo halabuurka allelic-ka Lanr1/AnMan ayaa loo doortay qaabaynta muujinta hiddaha difaaca ee kala duwan iyadoo la adeegsanayo laba hab oo is-waafaqsan: isku-dubaridka RNA-ga ee wax-soo-saarka sare leh iyo qiyaasta PCR-ga waqtiga-dhabta ah.
Baaritaanka set oo ah jeermiska NLL (N = 215) oo leh calaamado Lanr1 (Anseq3 iyo Anseq4) iyo AnMan (Anseq4) iyo AnMan (AnManM1) waxay muujiyeen in hal xariiq oo keliya (95726, oo u dhow Salamanca-b) ay kordhinayso allele-ka "iska caabbinta" dhammaan calaamadaha, halka "Joogitaanka allele-ka 'u nugul'" ay heshay saamiga dhammaan calaamadaha 158 (~73.5%) khadadka. Saddex iyo toban xariiq ayaa soo saaray laba allele "iska caabbinta" ee calaamadda Lanr1, 8 xariiqna waxay soo saareen allele "iska caabbinta" ee calaamadda Lanr1. allele-ka "iska caabbinta" ee calaamadda AnMan (Jadwalka Dheeraadka ah S1). Laba xariiq ayaa heterozygous u ahaa calaamadda Anseq3 iyo hal heterozygous oo loogu talagalay calaamadda AnManM1. 42 xariiq (19.5%) waxay qaadeen marxalado iska soo horjeeda oo ah alleles-ka Anseq3 iyo Anseq4, taasoo muujinaysa soo noqnoqosho sare oo isku-dhafan oo u dhexeeya labadan loci. Noocyada Anthracnose ee xaaladaha la xakameeyey (Jadwalka Dheeraadka ah ee S2) ayaa muujiyay kala duwanaansho ku jirta iska caabbinta noocyada hidde-sideyaasha ee la tijaabiyey, taas oo ka muuqatay darnaanta anthracnose. Kala duwanaanshaha dhibcaha celceliska waxay u dhexeeyeen 1.8 (dhexdhexaad u adkaysta) ilaa 6.9 (u adkaysta) kala duwanaanshaha miisaanka dhirtana waxay u dhexeeyeen 0.62 (u adkaysta) ilaa 4.45 g (u adkaysta). Waxaa jiray xiriir muhiim ah oo u dhexeeya qiimayaasha lagu arkay laba nuqul oo tijaabada ah (0.51 dhibcaha darnaanta cudurka, P = 0.00017 iyo 0.61 miisaanka dhirta, P < 0.0001) iyo sidoo kale labadan cabbir (− 0.59 iyo − 0.77, P < 0.0001). Waxaa jiray xiriir muhiim ah oo u dhexeeya qiimayaasha lagu arkay laba nuqul oo tijaabada ah (0.51 dhibcaha darnaanta cudurka, P = 0.00017 iyo 0.61 miisaanka dhirta, P < 0.0001) iyo sidoo kale labadan cabbir (− 0.59 iyo − 0.77, P < 0.0001). 0,513 блезни, P = 0,00017 и 0,61 для массы растения, P <0,0001), а также межdu этими двумя параметрами (-0,50 , 0,1) <0,59 и -0,1) 0,0001). Xiriir muhiim ah ayaa laga helay qiimaha lagu arkay laba ku celcelin oo tijaabada ah (0.51 dhibcaha darnaanta cudurka, P = 0.00017 iyo 0.61 miisaanka dhirta, P < 0.0001), iyo sidoo kale inta u dhaxaysa labadan cabbir (- 0.59 iyo -0.77, P < 0.0001) 0.0001).. 0.00017，植物重量为0.61，P <0.0001）以及这两个参数之间（- 0.59 和- 0.77，P <0.001)在 两 次重复 实验 中 观察 的 值 之间 存在 相关性, p = 0.00017, 植物 为 为 0.61和- 0.59 和 - 0.77, P <0.0001). Наблюдалась? 0,00017 iyo maassan растения 0,61, P <0,0001), iyo межdu эtymy дvyumya parametram (-0,59 iyo -0,0001) 0,77, P <0,0001. Waxaa jiray xiriir muhiim ah oo u dhexeeya qiimayaasha lagu arkay nuqulka (dhibcaha darnaanta cudurka 0.51, P = 0.00017 iyo miisaanka dhirta 0.61, P < 0.0001) iyo inta u dhaxaysa labadan cabbir (-0.59 iyo -0 .0001) 0.77, P<0.0001. ).Calaamadaha caadiga ah ee lagu arko dhirta u nugul waxaa ka mid ah leexashada iyo leexashada jirridda oo u eg qaab-dhismeedka "qaansada adhijirka", oo ay ku xigto nabarro qaab-dhismeed oo leh sporozoites oranji/casaan ah (Jaantuska Dheeraadka ah 1). Ku-biirinta Australiyaanka oo sidda hiddo-sidayaasha Lanr1 (83A:476 iyo Tanjil) iyo AnMan (Mandelup) waa kuwo iska caabiya dhexdhexaadka ah, 0.0331 iyo 0.0036). Khadadka qaarkood oo sidoo kale wata alleles Lanr1 iyo/ama AnMan "iska caabiya" waxay muujinayaan astaamaha cudurka.
Waxaa xiiso leh, dhowr xariiq oo NLL ah oo aan lahayn wax calaamad ah oo "iska caabiya" ayaa muujiyay heer sare oo iska caabbinta anthracnose ah (la barbar dhigi karo ama ka sarreeya noocyada Lanr1 ama AnMan), sida Boregine (qiimaha P < 0.0001 labada cabbir), Bojar (qiimaha P < 0.0001 dhibcaha iyo 0.001 miisaanka dhirta) iyo Dadweynaha B-549/79b (qiimaha P < 0.0001 dhibcaha iyo aan muhiim ahayn miisaanka). Waxaa xiiso leh, dhowr xariiq oo NLL ah oo aan lahayn wax calaamad ah oo "iska caabiya" ayaa muujiyay heer sare oo iska caabbinta anthracnose ah (la barbar dhigi karo ama ka sarreeya noocyada Lanr1 ama AnMan), sida Boregine (qiimaha P < 0.0001 labada cabbir), Bojar (qiimaha P < 0.0001 dhibcaha iyo 0.001 miisaanka dhirta) iyo Dadweynaha B-549/79b (qiimaha P < 0.0001 dhibcaha iyo aan muhiim ahayn miisaanka). Интересно, что несколько линий NLL, лишенных какого-либо устойчивости к антракнозу обоих параметров), Bojar (значение P <0,0001 для оценки и 0,001 для массы растения) популяции B-549/79b (значение P <0,0001 для оценки и незначимо для массы). Waxaa xiiso leh, dhowr xariiq oo NLL ah oo aan lahayn wax calaamad ah oo 'iska caabiya' ayaa muujiyay heer sare oo iska caabin ah oo loo qabo anthracnose (la barbar dhigi karo ama ka sarreeya noocyada Lanr1 ama AnMan), sida Boregine (qiimaha P < 0.0001 labada cabbir), Bojar (qiimaha P < 0.0001 qiimaynta iyo 0.001 miisaanka dhirta) iyo tirada dadka B-549/79b (qiimaha P < 0.0001 qiimaynta mana aha mid muhiim u ah miisaanka).有趣的是，一些缺乏任何“抗性”标记等位基因的NLL基因型相当或更高），例如Boregine（两个参数的P 值< 0.0001）、Bojar（P 值<得分为0.0001，植物重量为0.001）和种群B-549/79b（得分P 值< 0.0001，重量一） Waa wax xiiso leh in qaar ka mid ah nidaamyada NLL ee aan lahayn wax calaamado "antigenic" ah ay muujiyaan iska caabin sare oo toosan (oo u dhiganta hiddo-wadaha Lanr1 ama AnMan ama ka sareeya), sida Boregine (labada cabbir P < 0.0001), Bojar (qiimaha P < 0.0001, miisaanka dhirta 0.001) iyo nooca B-549/79b (qiimaha P < 0.0001, miisaanku ma aha mid muhiim ah). Интересно, что некоторые линии NLL. устойчивости к антракнозу (сравнимые или выше, чем у генотипов Lanr1 или AnMan), такие как Boregine Bojar (значение P <0,0001, масса растения 0,001) iyo популяция B-549/79b (оценка P-значение) <0,0001, масса незначительна). Waxaa xiiso leh, qaar ka mid ah khadadka NLL oo aan lahayn wax calaamado 'iska caabin ah' ah ayaa muujiyay heerar sare oo iska caabin ah oo anthracnose ah (la barbar dhigi karo ama ka sarreeya noocyada hiddo-wadaha Lanr1 ama AnMan), sida Boregine (qiimaha P ee labada cabbir <0.0001), Bojar (qiimaha P < 0.0001, miisaanka dhirta 0.001) iyo tirada dadka B-549/79b (qiimaha P < 0.0001, miisaanku ma aha mid muhiim ah).Ifafaaladdan waxay soo jeedinaysaa suurtagalnimada ilo cusub oo iska caabin hidde ah, iyadoo sharraxaysa la'aanta isku xirnaanta la arkay ee u dhaxaysa noocyada calaamadaha iyo noocyada cudurka (qiimaha P laga bilaabo ~0.42 ilaa ~0.98). Sidaa darteed, tijaabada Kolmogorov-Smirnov waxay muujisay in xogta ku saabsan iska caabbinta anthracnose ay si caadi ah ugu qaybsanayd dhibcaha (qiimaha P 0.25 iyo 0.11) iyo cufka dhirta (qiimaha P 0.47 iyo 0.55), taasoo soo jeedinaysa I waxay qiyaasaysaa in alleles ka badan Lanr1 iyo AnMan ay ku lug leeyihiin.
Iyada oo ku saleysan natiijooyinka baaritaanka iska caabbinta anthracnose, 4 sadar ayaa loo xushay falanqaynta qoraalka: 83A:476, Boregine, Mandelup, iyo Population 22660. Khadadkan waxaa dib loogu tijaabiyay iska caabbinta anthrax ee tijaabooyinka tallaalka iyadoo la adeegsanayo taxanaha RNA, iyadoo la raacayo inay la mid yihiin tijaabadii hore. Qiimaha dhibcaha waxay ahaayeen sidan soo socota: Boregin (1.71 ± 1.39), 83A: 476 (2.09 ± 1.38), Mandelup (3.82 ± 1.42) iyo dadweynaha 22660 (6.11 ± 1.29).
Hab-maamuuska Illumina NovaSeq 6000 wuxuu gaaray celcelis ahaan 40.5 lammaane Mread muunad kasta (29.7 ilaa 54.4 Mreads) (Jadwalka Dheeraadka ah S3). Dhibcaha isku-xidhka ee taxanaha tixraaca waxay u dhexeeyeen 75.5% ilaa 88.6%. Isku-xidhka celceliska xogta tirinta akhriska ee u dhaxaysa kala duwanaanshaha tijaabada ee u dhexeeya nuqullada bayoolojiga waxay u dhexeeyeen 0.812 ilaa 0.997 (celcelis ahaan 0.959). 35,170 hiddo-side oo la falanqeeyay, 2917 ayaa muujiyay wax muujin ah, 4785-kii hiddo-side ee kale waxaa lagu muujiyay heer aan la taaban karin (celceliska salka < 5). 35,170 hiddo-side oo la falanqeeyay, 2917 ayaa muujiyay wax muujin ah, 4785-kii hiddo-side ee kale waxaa lagu muujiyay heer aan la taaban karin (celceliska salka < 5). Иz 35 170 proanalizirovannыh genov 2917 iyo proyavlyaly эkspressi незначительном уровне (базовое среднее <5). 35,170-kii hiddo-side ee la falanqeeyay, 2917 ma muujin wax muujin ah, 4785-kii hiddo-side ee soo harayna waxaa lagu muujiyay heer aan la taaban karin (celceliska salka <5).在分析的35,170 个基因中，2917 个没有表达，其他4785 个基國5)35,170 Иz 35 170 proanalizirovannыh genov 2917 iyo Ostalnыe 4785 (базовое среднее значение <5). 35,170-kii hiddo-side ee la falanqeeyay, 2917 lama muujin, 4785-kii hiddo-side ee soo harayna waxay lahaayeen muujin aan la taaban karin (celceliska salka <5).Sidaa darteed, tirada hiddo-sidayaasha loo tixgeliyey in la muujiyay (celceliska salka ≥ 5) intii lagu jiray tijaabada waxay ahayd 27,468 (78.1%) (Jadwalka Dheeraadka ah S4).
Laga bilaabo markii ugu horreysay, dhammaan khadadka NLL waxay ka jawaabeen tallaalka C. lupini (nooca Col-08) iyagoo dib u barnaamijiyay qoraalka (Jadwalka 1), si kastaba ha ahaatee, waxaa la arkay kala duwanaansho muhiim ah oo u dhexeeya khadadka. Sidaa darteed, khadka iska caabbinta 83A:476 (oo sita hidda-wadaha Lanr1) wuxuu muujiyay dib-u-barnaamij muhiim ah oo ku saabsan qoraalka markii ugu horreysay (6 hpi) iyadoo 31-69-laab ay kor u kacday tirada hidda-wadaha kor iyo hoos u go'doonsan marka la barbar dhigo waqtiyada kale ee waqtigan. Intaa waxaa dheer, heerkan ugu sarreeya wuxuu ahaa mid gaaban, maadaama muujinta hiddo-wadeyaal yar oo keliya ay si weyn isu beddeleen markii labaad (12 hpi). Waxaa xiiso leh, Boregine, oo sidoo kale muujisay heer sare oo iska caabin ah oo ku saabsan tijaabada tallaalka, ma uusan marin dib-u-barnaamij qoraal oo aad u weyn intii lagu jiray tijaabada. Si kastaba ha ahaatee, tirada hiddo-wadaha si kala duwan loo muujiyey (DEG) waxay ahayd isku mid Boregine iyo 83A:476 12 HPI. Mandelup iyo dadweynaha 22660 labaduba waxay muujiyeen heerka ugu sarreeya ee DEG xilligii ugu dambeeyay (48 l/s), taasoo muujinaysa dib u dhac ku yimid jawaab celinta difaaca.
Sababtoo ah 83A:476 waxaa la mariyay dib-u-habeyn ballaaran oo ku saabsan qoraalka iyadoo laga jawaabayo C. lupini oo ah 6 HPI marka la barbar dhigo dhammaan khadadka kale, ~91% DEG-yada la arkay waqtigan waxay ahaayeen kuwo gaar u ah qabiilka (Jaantuska 1). Si kastaba ha ahaatee, waxaa jiray isku-dhacyo jawaabo hore oo u dhexeeya khadadka daraasadda, maadaama 68.5%, 50.9%, iyo 52.6% DEG ee Boregine, Mandelup, iyo dadweynaha 22660, siday u kala horreeyaan, ay isku dulsaarnaayeen kuwa laga helay 83A:476 waqtiyo gaar ah. Si kastaba ha ahaatee, DEG-yadani waxay xisaabiyeen oo keliya jajab yar (0.97–1.70%) dhammaan DEG-yada hadda la ogaado iyadoo la adeegsanayo 83A:476. Intaa waxaa dheer, 11 DEG oo ka yimid dhammaan xariiqyada ayaa isku mid ahaa waqtigan (Jadwallada Dheeraadka ah ee S4-S6), oo ay ku jiraan qaybaha caadiga ah ee jawaabaha difaaca dhirta: borotiinka wareejinta lipid (TanjilG_32225), ensaymka endoglucan-1,3-β-glucoside (TanjilG_23384), laba borotiin oo la kicin karo sida SAM22 (TanjilG_31528 iyo TanjilG_31531), borotiinka latex-ka aasaasiga ah (TanjilG_32352), iyo laba borotiin oo derbi unug oo qaab-dhismeed ah oo glycine-hodan ah (TanjilG_19701 iyo TanjilG_19702). Waxaa sidoo kale jiray isku-dhafnaan aad u sarreysa oo ku yimid jawaabaha qoraalka ah ee u dhexeeya 83A:476 iyo Boregine oo ah 24 HPI (wadarta 16-38% DEG) iyo inta u dhaxaysa Mandelup iyo Population 22660 oo ah 48 HPI (wadarta 14-20% DEG).
Jaantuska Venn oo muujinaya tirada hiddo-sideyaasha si kala duwan loo muujiyey (DEG) ee khadadka lupine-ka cidhiidhiga ah (NLL) ee lagu tallaalay Colletotrichum lupini (nooca Col-08 ee laga helay beeraha lupine ee Wierzhenice, Poland, 1999). Khadadka NLL ee la falanqeeyay waxay ahaayeen: 83A:476 (adkaysi, oo sidda allele-ka Lanr1), Boregine (adkaysi, asal hidde ah oo aan la garanayn), Mandelup (adkaysi dhexdhexaad ah, oo sidda allele-ka AnMan) iyo tirada dadka 22660 (aad u nugul). Soo gaabinta hpi waxay u taagan tahay saacado ka dib tallaalka. Qiimo eber ah ayaa laga saaray si loo fududeeyo garaafka.
Qaybaha hiddo-sidayaasha aadka loo muujiyey ee 6 hpi ayaa lagu falanqeeyay jiritaanka hidde-sidayaasha R ee canonical (Jadwalka Dheeraadka ah ee S7). Daraasaddani waxay shaaca ka qaadday soo-kicinta qoraalka ah ee hiddo-sidayaasha iska caabbinta cudurka ee caadiga ah oo leh hidde-sidayaasha NBS-LRR oo keliya 83A:476. Qaybtani waxay ka koobnayd hal hiddo-sidayaasha TIR-NBS-LRR (tanjilg_05042), shan hiddo-sidayaasha CC-NBS-LRR (tanjilg_06165, tanjilg_06162, tanjilg_22773, tanjilg_22640, iyo tanjilg_16162), iyo Afar NBS-LR, Tanjilg_16162), iyo Afar NBS-LRR (tanjilg_16162) iyo sidoo kale Afar NBS-Lrr (tanjilg_16162) iyo Afar NBS-LRR (TANJILG_16162). Dhammaan hiddo-sidayaashani waxay leeyihiin domains canonical ah oo loo habeeyay taxane la ilaaliyo. Marka laga soo tago hiddo-sidayaasha domainka NBS-LRR, dhowr kinases RLL ah ayaa la hawlgeliyay 6 hpi, kuwaas oo kala ah mid ku jira Boregine (TanjilG_19877), laba ku jira Mandelup (TanjilG_07141 iyo TanjilG_19877) iyo tirada dadka 22660 (TanjilG_09014 iyo TanjilG_10361) iyo laba ku jira 83A 27:476.
Hidde-sidayaasha oo si weyn isu beddelay muujinta ka dib markii lagu tallaalay C. lupini (nooca Col-08) ayaa lagu sameeyay falanqaynta kobcinta Gene Ontology (GO) (Jadwalka Dheeraadka ah ee S8). Ereyga habka bayoolojiga ee ugu badan ee aadka loo matalay wuxuu ahaa "Jawaabta difaaca GO:0006952" kaas oo ka soo muuqday 6 ka mid ah 16 isku-darka (waqtiga × xariiqda) oo leh muhiimad sare (qiimaha P < 0.001) (Jaantuska 2). Ereyga habka bayoolojiga ee ugu badan ee aadka loo matalay wuxuu ahaa "Jawaabta difaaca GO:0006952" kaas oo ka soo muuqday 6 ka mid ah 16 isku-darka (waqtiga × xariiqda) oo leh muhiimad sare (qiimaha P < 0.001) (Jaantuska 2). Наиболее часто чрезмерно представленным термином появлялся в 6 из 16 (время × линия) комбинаций с высокой значимостью (значение P <0,001) (рис. 2). Ereyga habka bayoolojiga ee ugu badan ee aadka loo matalay wuxuu ahaa 'Jawaabta difaaca GO:0006952', kaas oo ka soo muuqday 6 ka mid ah 16 isku-darka (waqtiga × jiilka) oo leh muhiimad sare (qiimaha P < 0.001) (Jaantuska 2).最常被过度代表的生物过程术语是“GO:0006952个中，具有高显着性（P 值< 0.001）（图2）。 Ereyga habka bayoolojiga ee ugu matalaadda badan waa "GO:0006952 jawaab celin difaac", kaas oo ka muuqda 6 ka mid ah 16ka isku-darka (时间×线), oo leh muhiimad sare (qiimaha P < 0.001) (图2). "GO: 0006952 Jawaabta Difaaca" , из 16 комбинаций (время × линия) с высокой значимостью (значение P <0,001) (рис. 2). Ereyga habka bayoolojiga ee ugu badan ee aadka loo matalay wuxuu ahaa 'GO:0006952 Difaaca Jawaabta', kaas oo ka soo muuqday 6 ka mid ah 16 isku-darka (waqtiga × xariiq) oo leh muhiimad sare (qiimaha P < 0.001) (Jaantuska 2).Ereygan waxaa lagu muujiyey laba jeer oo isku xiga 83A: 476 iyo Boregine (6 iyo 24 hpi) iyo hal mar oo isku xigta Mandelup iyo Population 22660 (12 iyo 6 hpi, siday u kala horreeyaan). Tani waa natiijo la filayo, taasoo muujinaysa jawaabta antifungal ee khadadka iska caabbinta. Intaa waxaa dheer, 83A:476 waxay ka jawaabtay C. lupini iyadoo si degdeg ah u kicinaysa hiddo-sideyaal la xiriira burburka oksaydhka oo lagu matalo ereyga "GO:0055114 redox process", taasoo muujinaysa jawaab difaac oo gaar ah, halka Boregine uu shaaca ka qaaday jawaabo difaac oo gaar ah, oo la xiriira ereyga 'GO'. :0006950 Jawaabta Cadaadiska". Dadweynaha 22660 waxay kicisay jawaabta iska caabbinta toosan ee ku lug leh dheef-shiid kiimikaadka labaad, iyagoo muujinaya tirada xad-dhaafka ah ee ereyada "GO:0016104 Geedi socodka biosynthesis-ka triterpene" iyo "GO:0006722 Geedi socodka dheef-shiid kiimikaadka triterpene" (labada ereyba waxay ka tirsan yihiin isla hidde-sideyaasha), iyadoo la tixgelinayo natiijooyinka falanqaynta kobcinta ereyga GO, xasilloonida falcelinta Mandelup waxay u dhaxaysay Boregine iyo Dadweynaha 22660. Intaa waxaa dheer, falcelinta hore 83A:476 (6 hpi) iyo falcelinta dib u dhaca Mandelup iyo Dadweynaha 22660 waxaa ka mid ah ereyga GO:0015979 'photosynthesis' iyo hababka kale ee bayoolojiga ee la xiriira.
Erayada ontology-ga hidde-sideyaasha bayoolojiga ee lagu xushay sharraxaadda hiddo-sideyaasha si kala duwan loo muujiyey inta lagu jiro jawaabaha qoraalka ee lupine-ka caleenta cidhiidhiga ah (NLL) ee lagu tallaalay lupine-ka anthrax (nooca Col-08 ee laga helay beeraha lupine ee Wierzhenice, Poland, sanadkii 1999) aad ayaa loo buunbuuniyay. Khadadka NLL ee la falanqeeyay waxay ahaayeen: 83A:476 (adkaysi, oo sita allele-ka homozygous Lanr1), Boregine (asalka hidde-sideyaasha adkaysi u leh, oo aan la garanayn), Mandelup (adkaysi dhexdhexaad ah, oo sita allele-ka homozygous AnMan) iyo dadweynaha 22660 (u nugul).
Maadaama daraasaddani ay ujeeddadeedu ahayd in la aqoonsado hiddo-sideyaasha gacan ka geysta iska caabbinta anthracnose, hiddo-sideyaasha loo qoondeeyay ereyada GO "GO: 0006952 Jawaabaha Difaaca" iyo "GO: 0055114 Hababka Redox" waxaa lagu falanqeeyay jaritaanno maadaama macnaha aasaasiga ah uu yahay ≥ 30 oo leh ugu yaraan hal sadar. × dhibic waqtiga oo isku daraysa qiimayaasha tirakoob ahaan muhiimka ah ee log2 (isbeddelka laablaabka). Tirada hiddo-sideyaasha buuxiya shuruudahan waxay ahayd 65 GO:0006952 iyo 524 GO:0055114.
83A:476 wuxuu shaaca ka qaaday laba meelood oo DEG ah oo lagu sharraxay ereyga GO:0006952, kan ugu horreeya 6 hidde-side halkii inji (64 hidde-side, nidaaminta kor iyo hoos) iyo kan labaad 24 hidde-side halkii inji (15 hidde-side, nidaaminta kor iyo hoos oo keliya). Boregine wuxuu sidoo kale muujiyay in GO:0006952 uu ugu sarreeyay isla waqtigaas, laakiin uu ka yar yahay DEG (11 iyo 8) iyo firfircoonida doorbidka leh. Mandeloop wuxuu muujiyay laba meelood oo GO ah:0006952 12 iyo 48 HPI, labaduba waxay wataan 12 hidde-side (kan ugu horreeya oo leh hidde-side firfircoon, kan labaadna wuxuu leeyahay hidde-sideyaal xakameynaya oo keliya), halka dadka 22660 ee 6 HPI (13 hidde-sideyaal) ay lahaayeen awood weyn oo ah nidaaminta kororka ugu sarreysa. Waa in la ogaadaa in 96.4% GO:0006952 DEG ee meelahan ugu sarreeya ay lahaayeen nooc isku mid ah oo jawaab celin ah (kor ama hoos), taasoo muujinaysa isku-dhafnaan weyn oo ku jirta jawaabaha difaaca inkastoo ay kala duwan yihiin tirada hidde-sideyaasha ku lug leh. Kooxda ugu weyn ee taxanaha ah ee la xiriira ereyga GO:0006952 waxay codsataa Borotiinka Farriinta ee la Xiriira Gaajada Cadaadiska-Isku-xirka 22 (SAM22-sida), kaas oo ka tirsan borotiinka la xiriira cudurka 'pathogenous-associated protein' (PR-10) iyo borotiinka asaasiga ah ee 'latex'. borotiinka la midka ah (MLP-sida) (Jaantuska 3). Labada kooxood waxay ku kala duwanaayeen dabeecadda muujinta iyo jihada jawaabta. Hiddo-sidayaasha codeeya borotiinada u eg SAM22 waxay muujiyeen soo-kicin joogto ah oo muhiim ah waqtiyadii hore (6 ama 12 hpi) waxayna guud ahaan ka jawaabeen dhammaadka tijaabada (48 hpi), halka borotiinada u eg MLP ay muujiyeen isku-dubarid 6 hpi. hpi. 83A:476 iyo Mandelup 48 hp/in, ku dhawaad ​​​​dhammaan dhibcaha xogta kale ma aysan jawaabin. Intaa waxaa dheer, kala duwanaanshaha astaamaha muujinta ee hiddo-sidayaasha borotiinka u eg SAM22 waxay raaceen kala duwanaansho la arkay oo ku saabsan iska caabbinta anthracnose, maadaama khadadka iska caabbinta badan ay lahaayeen dhibco waqti oo badan oo si weyn u kiciya hiddo-sidayaashan marka loo eego hiddo-sidayaasha u nugul. Hiddo-side kale oo PR-10 u eg LlR18A/B ayaa muujiyay qaab muujin aad u la mid ah hiddo-side borotiin u eg SAM22.
Qaybaha ugu muhiimsan ee ereyga habka bayoolojiga "GO:0006952 Jawaabta Difaaca" iyo qaababka muujinta hiddo-wadaha musharixiinta ah ee alleles-ka Lanr1 iyo AnMan ayaa la aqoonsaday. Miisaanka Log2 wuxuu matalaa qiimayaasha log2 (isbeddel laablaab ah) inta u dhaxaysa tallaalka (Colletotrichum lupini, nooca Col-08, laga helay beeraha lupine, Wizhenica, Poland, 1999) iyo dhirta xakamaynta (sham-inoculated) isla waqtigaas. Khadadka lupine ee caleenta cidhiidhiga ah ee soo socda ayaa la falanqeeyay: 83A:476 (adkaysi, oo sidda allele Lanr1 homozygous), Boregine (adkaysi, asal hidde ah oo aan la garanayn), Mandelup (adkaysi dhexdhexaad ah, oo sidda allele AnMan homozygous), iyo Population 22660 (u nugul).
Intaa waxaa dheer, astaamaha muujinta ee hiddo-sidayaasha musharraxiinta RNA-seq ee Lanr1 (TanjilG_05042) iyo AnMan (TanjilG_12861) ayaa la qiimeeyay (Jaantuska 3). Hiddo-sidayaasha TanjilG_05042 waxay muujiyeen jawaab celin muhiim ah (dhaqdhaqaaq) 83A:476 oo keliya markii ugu horreysay (6 hpi), halka TanjilG_12861 uu muhiim u ahaa Mandeloop oo keliya laba dhibcood oo waqti ah: 6 hpi (hoos u dhigista) iyo 24 hpi (6 hpi). Iyadoo.). la hagaajin karo) ).
Hidde-sidayaasha ugu badan ee lagu cabiray ereyga "habka redox" ee GO:0055114 waxay ahaayeen hiddo-sidayaasha codeeya borotiinada cytochrome P450 iyo peroxidase (Jaantuska 4). Muunadaha laga soocay 83A:476 ee 6 HPI, qiimaha ugu badan ama ugu yar ee log2 (isbeddelka laablaabka) (86.6% hiddo-sidayaasha) guud ahaan waxaa lagu arkay inta u dhaxaysa dhirta la tallaalay iyo kuwa la xakameeyey, taasoo muujinaysa jawaabta sare ee hiddo-sidayaashan ee galmada la tallaalay. 83A:476 waxay muujisay GO-ga ugu muhiimsan: 0055114 DEG ee 6 hpi (503 hiddo-sidayaasha), halka inta kale ee xariiqyada ay yihiin 48 hpi (Boregine, 31 hiddo-sidayaasha; Mandelup, 85 hiddo-sidayaasha; iyo Population 22660, 78 hiddo-sidayaasha)). Inta badan hiddo-sidayaasha qoyska GO:0055114, laba nooc oo jawaabo ah oo ku saabsan tallaalka (kicinta iyo joojinta) ayaa la arkay. Waxaa xiiso leh, ilaa 97.6% DEG-yada loo aqoonsaday ereyga GO: 0055114 ee Mandelupe oo ah 48 hp Indha-indheyntani waxay soo jeedinaysaa, inkastoo miisaanku aad u yar yahay (tusaale ahaan, tirada hiddo-wadaha redox ee isbeddelay, 85 marka loo eego 503), qaabka jawaabaha dib u dhaca ee transcription-ka ee mandeloup ilaa anthracnose waxay la mid tahay jawaab celinta hore ee 83A:476. Boregine iyo Population 22660, isku-darkani wuu ka hooseeyaa 51.6% iyo 75.6%, siday u kala horreeyaan.
Qaababka muujinta qaybaha ugu muhiimsan ee ereyga habka bayoolojiga "GO:0055114 Redox process" ayaa la shaaciyay. Miisaanka Log2 wuxuu matalaa qiimayaasha log2 (isbeddel laablaab ah) inta u dhaxaysa tallaalka (Colletotrichum lupini, nooca Col-08, laga helay beeraha lupine, Wizhenica, Poland, 1999) iyo dhirta xakamaynta (sham-inoculated) isla waqtigaas. Khadadka lupine ee caleenta cidhiidhiga ah ee soo socda ayaa la falanqeeyay: 83A:476 (adkaysi, oo sidda allele Lanr1 homozygous), Boregine (adkaysi, asal hidde ah oo aan la garanayn), Mandelup (adkaysi dhexdhexaad ah, oo sidda allele AnMan homozygous), iyo Population 22660 (u nugul).
83A:476 Jawaabaha qoraalka ah ee tallaalka C. lupini (nooca Col-08) waxa kale oo ku jiray aamusinta isku-dhafan ee hiddo-sideyaasha loo aaneeyay ereyga "sawir-qaadis" GO:0015979 iyo habab kale oo bayooloji ah oo la xiriira (Jaantuska 5). GO:0015979 DEG-gan waxa uu ka koobnaa 105 hiddo-side oo si weyn loogu cadaadiyay 6 hpi at 83A:476. Qaybtan, 37 hiddo-side ayaa sidoo kale hoos loogu dhigay Mandelup at 48 HPI iyo 35 at isla waqtigaas ee dadka 22660, oo ay ku jiraan 19 DEG-side oo caadi u ah labada nooc ee hiddo-sideyaasha. Ma jiraan DEG-sideyaal la xiriira ereyga GO: 0015979 oo si weyn loogu hawlgeliyay isku-darka kasta (khadka x waqtiga).
Qaababka muujinta qaybaha ugu muhiimsan ee ereyga habka bayoolojiga "GO:0015979 Photosynthesis" ayaa la shaaciyay. Miisaanka Log2 wuxuu matalaa qiimayaasha log2 (isbeddel laablaab ah) inta u dhaxaysa tallaalka (Colletotrichum lupini, nooca Col-08, laga helay beeraha lupine, Wizhenica, Poland, 1999) iyo dhirta xakamaynta (sham-inoculated) isla waqtigaas. Khadadka lupine ee caleenta cidhiidhiga ah ee soo socda ayaa la falanqeeyay: 83A:476 (adkaysi, oo sidda allele Lanr1 homozygous), Boregine (adkaysi, asal hidde ah oo aan la garanayn), Mandelup (adkaysi dhexdhexaad ah, oo sidda allele AnMan homozygous), iyo Population 22660 (u nugul).
Iyada oo lagu saleynayo natiijooyinka falanqaynta kala duwanaanshaha muujinta iyo sida la filayo inay ku lug leedahay jawaabaha difaaca ee ka dhanka ah fangaska cudurada keena, qaybtan toddobada hiddo-side ayaa loo xushay in lagu qiyaaso astaamaha muujinta iyadoo loo marayo PCR-ga waqtiga-dhabta ah (Jadwalka Dheeraadka ah ee S9).
Hidde-sidayaasha borotiinka ee TanjilG_10657 ayaa si weyn loogu kiciyay dhammaan khadadka la bartay iyo dhibcaha waqtiga marka la barbar dhigo dhirta xakamaynta (ee la midka ah) (Jadwallada Dheeraadka ah S10, S11). Intaa waxaa dheer, muuqaalka muujinta ee TanjilG_10657 wuxuu muujiyay isbeddel sii kordhaya inta lagu guda jiro tijaabada dhammaan khadadka. Dadweynaha 22660 waxay muujiyeen xasaasiyadda ugu sarreysa ee TanjilG_10657 ee tallaalka iyadoo la adeegsanayo firfircooni 114-laab ah iyo heerka muujinta qaraabada ugu sarreysa (4.4 ± 0.4) 24 HPI (Jaantuska 6a). Hidde-sidayaasha borotiinka PR10 LlR18A TanjilG_27015 ayaa sidoo kale muujiyay firfircooni dhammaan khadadka iyo dhibcaha waqtiga, iyadoo muhiimad tirakoob leh ay leedahay inta badan goobaha xogta (Jaantuska 6b). Sida TanjilG_10657 oo kale, heerka muujinta qaraabada ugu sarreysa ee TanjilG_27015 ayaa lagu arkay 22660 oo qof oo la tallaalay 24 HPI (19.5 ± 2.4). Hiddo-wadaha aashitada ee TanjilG_04706 ayaa si weyn kor ugu kacay dhammaan khadadka iyo waqtiyada oo dhan marka laga reebo Boregine 6 hpi (Jaantuska 6c). Waxaa si xooggan u kiciyay markii ugu horreysay (6 HPI) 83A:476 (10.5 jeer) waxaana si dhexdhexaad ah u kordhay khadadka kale (6.6-7.5 jeer). Intii lagu jiray tijaabada, muujinta TanjilG_04706 waxay ku sii jirtay heerar la mid ah 83A:476 iyo Boregine, halka Mandelup iyo Population 22660 ay si weyn u korodhay, iyadoo gaartay qiimayaal aad u sarreeya (5.9 ± 1.5 iyo 6.2 ± 1.5, siday u kala horreeyaan). Hiddo-wadaha u eg endoglucan-1,3-β-glucosidase ee TanjilG_23384 ayaa muujiyay firfircooni sare labadii dhibcood ee ugu horreeyay (6 iyo 12 hpi) dhammaan khadadka marka laga reebo tirada dadka 22660 (Jaantuska 6d). Heerarka muujinta qaraabada ugu sarreeya ee TanjilG_23384 ayaa lagu arkay barta labaad (12 hpi) ee Mandelup (2.7 ± 0.3) iyo 83A:476 (1.5 ± 0.1). 24 HPI, muujinta TanjilG_23384 waxay ahayd mid aad u hooseysa dhammaan khadadka la bartay (laga bilaabo 0.04 ± 0.009 ilaa 0.44 ± 0.12).
Astaamaha muujinta hiddo-sidayaasha la doortay (ag) oo lagu muujiyay PCR tiro ahaan. Tirooyinka 6, 12 iyo 24 waxay matalaan saacado ka dib tallaalka. Hiddo-sidayaasha LanDExH7 iyo LanTUB6 waxaa loo isticmaalay caadiyaynta halka LanTUB6 loo isticmaalay habaynta taxanaha dhexdooda. Baararka khaladaadka waxay matalaan leexashada caadiga ah ee ku salaysan saddex nuqul oo bayooloji ah, mid walbana waa celceliska saddex nuqul oo farsamo ah. Muhiimadda tirakoobka ee kala duwanaanshaha heerarka muujinta ee u dhexeeya tallaalka la tallaalay (Colletotrichum lupini, nooca Col-08, oo laga helay 1999 beerta lupin ee Wierzenica, Poland) iyo dhirta xakamaynta (ku-tallaalidda lagu sameeyay) waxaa lagu calaamadeeyay dhibcaha xogta ee kor ku xusan (*Qiimaha P < 0.05, **Qiimaha P ≤ 0.01, ***Qiimaha P ≤ 0.001). Muhiimadda tirakoobka ee kala duwanaanshaha heerarka muujinta ee u dhexeeya tallaalka la tallaalay (Colletotrichum lupini, nooca Col-08, oo laga helay 1999 beerta lupin ee Wierzenica, Poland) iyo dhirta xakamaynta (ku-tallaalidda lagu sameeyay) waxaa lagu calaamadeeyay dhibcaha xogta ee kor ku xusan (*Qiimaha P < 0.05, **Qiimaha P ≤ 0.01, ***Qiimaha P ≤ 0.001). Статическая значимость различий v urrovnyah эkspressi mezhdu iyokulyirovannыmy (Colletotrichum lupini, shtamm Col-08, поля люпина в Верженице, Польша) iyo контрольным 0,05, **значение P ≤ 0,01, ***значение P ≤ 0,001). Kala duwanaanshaha tirakoob ahaan muhiimka ah ee heerarka muujinta ee u dhexeeya tallaalka (Colletotrichum lupini, nooca Col-08, oo laga helay 1999 beerta lupine ee Wierzhenice, Poland) iyo dhirta xakamaynta (sham-inoculated) ayaa lagu xusay dhibcaha xogta kor ku xusan (*Qiimaha P < 0.05, **Qiimaha P ≤ 0.01, ***Qiimaha P ≤ 0.001).接种（Colletotrichum lupini, Col-08株，1999年从波兰Wierzenica的羽扇豆田获得）和对照（模拟接种）植物之间表达水平差异的统计学显着性标记在数捉捉丹上在数捹丹伸0.05, ** P 值≤ 0.01, ***P 值≤ 0.001).接种 （colletotrichum lupini, color-08 株, 1999 年 波兰 波兰 wierzenica水平 差异 的 统计学 Sida laga soo xigtay wargeyska caanka ah, Colletotrichum lupini, Col-08, iwm. люпина в Верженице, Польша, в 1999 г.) iyo контрольныmy значение P <0,05, ** P-значение ≤ 0,01, ***P-значение ≤ 0,001). Kala duwanaanshaha tirakoob ahaan muhiimka ah ee heerarka muujinta ee u dhexeeya tallaalka (Colletotrichum lupini, nooca Col-08, laga helay beeraha lupine ee Verzhenice, Poland, sanadkii 1999) iyo dhirta xakamaynta (sham-inoculated) ayaa lagu xusay dhibcaha xogta kor ku xusan (*Qiimaha P < 0.05 , **Qiimaha P ≤ 0.01, ***Qiimaha P ≤ 0.001).Khadadka NLL ee la falanqeeyay waxay ahaayeen: 83A:476 (adkaysi u leh, oo sidda allele-ka homozygous Lanr1), Mandelup (adkaysi dhexdhexaad ah u leh, oo sidda allele-ka homozygous AnMan), Boregine (asal hidde-side oo adkaysi u leh, oo aan la garanayn) iyo dadweynaha 22660 (u nugul).
Hidde-sidayaasha musharixiinta ah ee TanjilG_05042 ee goobta Lanr1 waxay muujiyeen qaab muujin oo aad uga duwan astaamaha laga helay daraasadaha RNA-seq (Jaantuska 6e). Dhaqdhaqaaq muhiim ah oo hidda-wadahan ah ayaa lagu arkay Mandelup iyo 22660 qof (ilaa 39.7 iyo 11.7 jeer, siday u kala horreeyaan), taasoo keentay heerar muujin oo aad u sarreeya (ilaa 1.4 ± 0.14 iyo 7.2 ± 1.3, siday u kala horreeyaan). 83A:476 sidoo kale waxay muujisay kor u kaca hidda-wadaha TanjilG_05042 (ilaa 3.8-laab), si kastaba ha ahaatee, heerarka muujinta qaraabada ah ee la gaaray (0.044 ± 0.002) waxay ka badnaayeen 30-laab ka hooseeya kuwa lagu arkay Mandelup iyo 22660 qof. Falanqaynta qPCR waxay muujisay kala duwanaansho muhiim ah oo ku saabsan heerarka muujinta ee u dhexeeya noocyada hiddo-sidayaasha ee noocyada tallaalka ee la tallaalay (la xakameeyey), iyagoo gaaray farqi 58-laab ah oo u dhexeeya dadka 22660 iyo 83A:476, iyo sidoo kale dadka 22660 iyo 22660. Farqi laba-laab ah ayaa laga gaaray Boregine iyo Mandalup.
Hidde-sidayaasha musharixiinta ah ee goobta AnMan, TanjilG_12861, ayaa la hawlgeliyay iyadoo laga jawaabayo tallaalka 83A:476 iyo Mandelup, waxay ahaayeen kuwo dhexdhexaad ah dadka 22660, waxaana hoos loogu dhigay Boregine (Jaantuska 6f). Muujinta qaraabada ah ee hidda-wadaha TanjilG_12861 ayaa ahayd tii ugu sarreysay ee tallaalka 83A: 476 (0.14±0.01). Hidde-sidayaasha borotiinka shoogga kulaylka ee fasalka I ee TanjilG_05080 HSP17.4 waxay muujiyeen heerar hoose oo muujinta qaraabada ah dhammaan noocyada la bartay iyo dhibcaha waqtiga (Jaantuska 6g). Qiimaha ugu sarreeya waxaa lagu arkay 24 HPI dadka 22660 (0.14 ± 0.02, koror siddeed laab ah oo ka dhashay tallaalka).
Isbarbardhigga astaamaha muujinta hidda-wadaha (Jaantuska 7) wuxuu muujiyay xiriir sare oo u dhexeeya TanjilG_10657 iyo afar hiddo-wade oo kale: TanjilG_27015 (r = 0.89), TanjilG_05080 (r = 0.85), TanjilG_05042 (r = 0.80), iyo TanjilG_04706 (r = 0.79). Natiijooyinka noocan oo kale ah waxay muujin karaan isku-dubaridka hidda-wadahaan inta lagu jiro jawaabaha difaaca. Hiddo-wadeyaasha TanjilG_12861 iyo TanjilG_23384 waxay muujiyeen astaamo muujin oo kala duwan oo leh qiimayaal isku-dhafan oo Pearson ah oo hooseeya (laga bilaabo 0.08 ilaa 0.43 iyo -0.19 ilaa 0.28, siday u kala horreeyaan) marka la barbar dhigo hiddo-wadeyaasha kale.
Isku xirnaanta u dhaxaysa astaamaha muujinta hidda-wadaha waxaa lagu ogaaday iyadoo la adeegsanayo PCR tiro ahaan. Khadadka lupine ee caleenta cidhiidhiga ah ee soo socda ayaa la falanqeeyay: 83A:476 (adkaysi u leh, oo sidda allele-ka homozygous Lanr1), Mandelup (adkaysi dhexdhexaad ah u leh, oo sidda allele-ka homozygous AnMan), Boregine (adag, asal hidde ah oo aan la garanayn), iyo Population 22660 (u nugul). Saddex dhibcood oo waqti ayaa la xisaabiyay (6, 12 iyo 24 saacadood ka dib tallaalka), oo ay ku jiraan tallaal (Colletotrichum lupini, nooca Col-08, oo laga helay beeraha lupine ee Wierzhenice, Poland, sanadkii 1999) iyo dhirta xakamaynta (sham-ku tallaalay). Miisaanka ayaa muujinaya qiimaha isku-xidhka Pearson.
Iyada oo lagu saleynayo xogta laga helay 6 faras koronto halkii inji, WGCNA waxaa lagu sameeyay 9981 DEG oo la aqoonsaday iyadoo la barbar dhigayo dhirta la tallaalay iyo kuwa la xakameeyay si diiradda loo saaro jawaabaha difaaca hore (Jadwalka Dheeraadka ah ee S12). Laba iyo labaatan qaybood oo hiddo-side ah (kooxo) ayaa la helay iyagoo leh astaamo muujinaya isku xiran (togan ama taban) oo u dhexeeya noocyada hiddo-sideyaasha iyo noocyada tijaabada. Celcelis ahaan, heerarka muujinta hidda-wadaha ayaa hoos u dhacayay si waafaqsan 83A:476 > Mandelup > Boregine > Dadweynaha 22660 (si kastaba ha ahaatee, labada noocba, isbeddelkani wuu ka xoog badnaa dhirta xakamaynta). Celcelis ahaan, heerarka muujinta hidda-wadaha ayaa hoos u dhacayay si waafaqsan 83A:476 > Mandelup > Boregine > Dadweynaha 22660 (si kastaba ha ahaatee, labada noocba, isbeddelkani wuu ka xoog badnaa dhirta xakamaynta). 83A:476 > Mandelup > Boregine > Dadka 22660 (dadka 22661 была сильнее у контрольных растений). Celcelis ahaan, heerarka muujinta hidda-wadaha ayaa hoos u dhacay sida ku xusan 83A:476 > Mandelup > Boregine > Dadweynaha 22660 (si kastaba ha ahaatee, labada noocba, isbeddelkani wuu ka xoog badnaa dhirta xakamaynta).平均而言，基因表达水平按83A:476 > Mandelup > Boregine > Dadka 22660 .平均 而 言 ， 基因 水平 按 按 83a: 476> mandelup> boregine> tirada dadka 22660 .中 更）                                                                        83A: 476 > Mandelup > Boregine > Dadka 22660 сильнее у контрольных растений). Celcelis ahaan, heerarka muujinta hidda-wadaha ayaa hoos u dhacay taxanaha 83A:476 > Mandelup > Boregine > Dadweynaha 22660 (si kastaba ha ahaatee, labada noocba, isbeddelkani wuxuu ka xoog badnaa dhirta xakamaynta).Tallaalku wuxuu keenay kor u kaca muujinta hidda-wadaha, gaar ahaan qaybaha 18, 19, 14, 6 iyo 1 (sida hoos u dhac ku yimid saamaynta), nidaaminta taban (tusaale qaybaha 9 iyo 20) ama saameyn dhexdhexaad ah (tusaale qaybaha 11, 22, 8 iyo 13). Falanqaynta kobcinta muddada GO (Jadwalka Dheeraadka ah S13) ayaa shaaca ka qaaday "GO: 0006952 Jawaabaha Ilaalinta" ee qaybta la tallaalay (18) oo leh firfircooni ugu badan, oo ay ku jiraan hiddo-sideyaal ay falanqeeyeen qPCR (TanjilG_04706, TanjilG_23384, TanjilG_10657 iyo TanjilG_27015), iyo sidoo kale qaybo badan oo sawir-qaadis ah oo Inoculate ah oo aad loo xakameeyey (9). Isu-ururinta Module 18 (Jaantuska 8) waxaa loo aqoonsaday hidda-wadaha TanjilG_26536 oo qeexaya borotiinka LlR18B ee PR-10-sida, isu-ururinta Module 9na waxaa loo aqoonsaday hidda-wadaha TanjilG_28955 oo qeexaya borotiinka photosystem II PsbQ. Hidda-wadaha iska caabbinta anthracnose ee Lanr1, TanjilG_05042, ayaa laga helay module 22 (Jaantuska 9) waxaana lala xiriiriyaa ereyada "GO:0044260 Hababka dheef-shiid kiimikaadka macromolecular-ka unugyada" iyo "GO:0006355 Xeer-dejinta qoraalka, qaabaynta DNA-da" oo sita xarunta TanjilG_01212. hidda-wadaha wuxuu codeeyaa qodobka qoraalka cadaadiska kulaylka A-4a (HSFA4a).
Falanqaynta shabakadda ee miisaanka leh ee muujinta isku-dhafka hiddaha ee modules-yada leh ereyada habka bayoolojiga ee aadka loo matalay "GO: 0006952 Jawaabaha Difaaca". Ligeration-ka ayaa la fududeeyay si loo muujiyo afarta hidde-side ee lagu falanqeeyay qPCR (TanjilG_04706, TanjilG_23384, TanjilG_10657 iyo TanjilG_27015).
Falanqaynta shabakadda ee miisaanka leh ee muujinta hidda-wadaha ee module leh erey geedi socod bayooloji oo aad loo matalay "GO: 0006355: Nidaaminta qoraalka, qaabaynta DNA-da" iyo sida hidda-wadaha iska caabbinta anthracnose ee Lanr1 TanjilG_05042. Laabista ayaa la fududeeyay si loo kala saaro hidda-wadaha TanjilG_05042 iyo hidda-wadaha TanjilG_01212 ee dhexe.
Baaritaanka iska caabbinta Anthracnose ee laga soo ururiyay Australia ayaa muujiyay in inta badan noocyada hore ee la sii daayay ay u nugul yihiin; Kalya, Coromup iyo Mandelup waxaa lagu tilmaamay inay yihiin kuwo iska caabbinta dhexdhexaadka ah, halka Wonga, Tanjil iyo 83A:476 lagu tilmaamay inay aad u adkaysi badan yihiin26,27,31. waxay lahaayeen isla allele iska caabbinta, oo loo qoondeeyay Lanr1, Coromup iyo Mandelupna waxay lahaayeen allele ka duwan, oo loo qoondeeyay AnMan10, 26, 39, halka Kalya ay gudbisay allele ka duwan., Lanr2. Baaritaanka iska caabbinta anthracnose ee Jarmalka wuxuu keenay in la aqoonsado xariiq iska caabbinta Bo7212 oo leh allele musharax ah oo aan ahayn Lanr1, oo loo qoondeeyay LanrBo36.
Daraasaddeennu waxay muujisay soo noqnoqosho aad u hooseysa (qiyaastii 6%) oo ah allele-ka Lanr1 ee ku jira germplasm-ka la tijaabiyay. Aragtidani waxay la jaanqaadaysaa natiijooyinka baaritaanka germplasm-ka Bariga Yurub iyadoo la adeegsanayo calaamadaha Anseq3 iyo Anseq4, taas oo muujisay in allele-ka Lanr1 uu ku jiro laba xariiq oo Belarusiyaan ah oo keliya. Tani waxay soo jeedinaysaa in allele-ka Lanr1 aan weli si weyn loo isticmaalin barnaamijyada taranka maxalliga ah, si ka duwan Australia, halkaas oo ay ka mid tahay allele-ka muhiimka ah ee taranka lagu caawiyo calaamadda. Tani waxay sabab u noqon kartaa heerka iska caabinta hooseeya ee allele-ka Lanr1 ee xaaladaha beeraha Yurub marka la barbar dhigo warbixinta Australia. Intaa waxaa dheer, daraasadaha anthracnose ee meelaha roobku badan yahay ee Australia waxay muujiyeen in jawaabaha iska caabinta ee ay dhexdhexaadiso allele-ka Lanr1 laga yaabo inaysan waxtar u lahayn xaaladaha cimilada ee door bidaaya koritaanka iyo horumarka degdegga ah ee pathogen-ka19,42. Dhab ahaantii, daraasaddan hadda jirta, qaar ka mid ah astaamaha anthracnose ayaa sidoo kale lagu arkay genotypes-ka sita allele-ka Lanr1, taasoo soo jeedinaysa in iska caabintu ay baaba'i karto xaaladaha ugu wanaagsan ee horumarinta C. lupini. Intaa waxaa dheer, fasiraado been abuur ah oo togan oo ku saabsan jiritaanka calaamadaha Anseq3 iyo Anseq4, kuwaas oo qiyaastii ah 1 cM laga bilaabo goobta Lanr1, ayaa suurtagal ah 28,30,43.
Daraasaddeennu waxay muujisay in 83A:476, oo sidda allele-ka Lanr1, ay ka jawaabtay tallaalka C. lupini iyadoo dib-u-habeyn ballaaran lagu sameeyay transcriptome-ka waqtiga ugu horreeya ee la falanqeeyay (6 hpi), halka Mandelup, oo sidda allele-ka AnMan, jawaabaha transcriptomic-ka la arkay waqti dheer ka dib. (laga bilaabo 24 ilaa 48 hp). Kala duwanaanshahan ku-meel-gaarka ah ee jawaabaha difaaca waxay la xiriiraan kala duwanaanshaha calaamadaha cudurka, iyagoo muujinaya muhiimadda ay leedahay aqoonsiga cudur-sidaha hore si loogu guuleysto jawaab celinta iska caabbinta. Si loo qaadsiiyo unugyada dhirta, unugyada anthrax waa inay maraan dhowr marxaladood oo horumarineed oo ku yaal dusha sare ee martida loo yahay, oo ay ku jiraan biqilka, qaybinta unugyada, iyo sameynta appressorium. Ku-darka waa qaab-dhismeed caabuq leh oo ku dheggan dusha sare ee martida loo yahay oo fududeeya gelinta unugyada martida loo yahay. Sidaa darteed, unugyada C. gloeosporioides ee laga soo saaray digirta ayaa muujiyay qaybtii ugu horreysay ee xudunta ka dib 75-90 daqiiqo oo la beeray, sameynta tuubo jeermis ah ka dib 90-120 daqiiqo, iyo xakamaynta ka dib 4 saacadood 45. Mango C. gloeosporioides waxay muujisay in ka badan 40% biqilka conidial ka dib 3 saacadood oo la beeray iyo qiyaastii 20% sameynta abresors ka dib 4 saacadood. Hidde-sidaha CAP20 ee C. gloeosporioides ee la xiriira fayraska ayaa muujiyay firfircooni qoraal ah oo ku jirta conidia-da sameysa epiphyte ka dib 3.5 saacadood oo lagu beeray shamaca dusha sare ee avokado oo leh uruurin sare oo borotiin CAP20 ah ka dib 4 saacadood iyo 46 daqiiqo. Sidoo kale, dhaqdhaqaaqa hiddo-sidayaasha biosynthesis ee melanin ee C. trifolii waxaa la kiciyay inta lagu jiro 2 saacadood oo lagu beeray ka dibna waxaa la sameeyay appressorium ka dib 1 saac. Daraasadaha unugyada caleenta ayaa muujiyay in strawberries-ka lagu tallaalay C. acutatum ay leeyihiin xakamaynta koowaad ee 8 hpi, halka yaanyada lagu tallaalay C. coccodes ay leeyihiin xakamaynta koowaad ee 4 hpi48,49. inta badan waxay la jaan qaadayaan cabbirka waqtiga habka faafa ee Colletotrichum spp.. Jawaabaha difaaca degdegga ah ee 83A:476 waxay soo jeedinayaan ku lug lahaanshaha iska caabbinta dhirta iyo hiddo-sidayaasha difaaca jirka ee saamaynta ku leh (ETI) ee xariiqdan, halka jawaabaha dib u dhaca ee Mandelup ay taageeraan mala-awaalka difaaca jirka ee la xiriira qaabka molecular-ka ee micro-la xiriira (MTI) 50. Jawaabaha hore ee 83A: 476 iyo Mandelup. Isku-darka qayb ahaan u dhexeeya hiddo-sidayaasha kor u kaca ama hoos u dhaca ee jawaabta dib u dhaca ayaa sidoo kale taageeraya fikraddan, maadaama ETI badanaa loo arko inay tahay jawaab degdeg ah oo la xoojiyay oo MTI ah oo ku dhammaada dhimashada unugyada ee la barnaamijiyay goobta cudurka, oo loo yaqaan shoogga anaphylactic 51,52.
Inta badan hiddo-sidayaasha loo aaneeyay ereyga aadka loo matalay ee Gene Ontology GO:0006952 "Jawaabta Difaaca" waa 11 isku-dhafan oo ka mid ah borotiinka soonka ee walaaca dhaliya ee 22 (oo la mid ah SAM22) iyo toddobada borotiin ee ugu waaweyn ee latex-ka u eg (MLPs). - borotiinno 31, 34, 43 iyo 423 waxay muujiyeen isku-mid ahaanshaha taxanaha. Hiddo-sidayaasha u eg SAM22 waxay muujiyeen firfircooni muhiim ah oo socotay muddo dheer, taasoo muujinaysa heerarka sii kordhaya ee iska caabbinta anthracnose (83A:476 iyo Boregine). Si kastaba ha ahaatee, hiddo-sidayaasha u eg MLP waxaa hoos loo dhigay oo keliya khadadka sita allele iska caabbinta musharaxa (83A:476/Lanr1 at 6 hpi iyo Mandelup/AnMan at 24 hpi). Waa in la ogaadaa in dhammaan isku-dhafannada SAM22-ka ee la aqoonsaday ay ka soo jeedaan koox hidde-side ah oo qiyaastii 105 kb ah, halka hiddo-sidayaasha u eg MLP ay ka soo jeedaan gobollo kala duwan oo ka mid ah hiddo-sidayaasha. Dadaallada isku-dubaridka ah ee hiddo-sidayaasha SAM22-ka oo kale ah ayaa sidoo kale laga helay daraasaddeennii hore ee ku saabsan iska caabbinta NLL ee tallaalka Diaporthetoxica, taasoo soo jeedinaysa inay ku lug leeyihiin qaybaha toosan ee jawaabta difaaca. Gunaanadkan waxaa sidoo kale taageeraya warbixinnada ku saabsan jawaab celin togan oo hiddo-sidayaasha SAM22-ka ah oo ku saabsan dhaawaca ama daaweynta salicylic acid, fangaska kiciya, ama hydrogen peroxide.
Hidde-sidayaasha MLP-ka ayaa la muujiyay inay ka jawaabaan walaacyo kala duwan oo ku saabsan abiotic iyo biotic, oo ay ku jiraan caabuqyada bakteeriyada, fayraska iyo cudurada faafa ee noocyada dhirta badan55. Tilmaamaha jawaab celinta isdhexgalka qaarkood ee u dhexeeya dhirta iyo cudurada faafa waxay u dhexeeyeen si xooggan oo sii kordhaya (tusaale ahaan, inta lagu jiro cudurka suufka ee Verticillium dahliae) ilaa si weyn hoos ugu dhaca (tusaale ahaan, ka dib markii uu ku dhaco geedka tufaaxa ee Alternaria spp.)56,57. Hoos u dhac weyn oo ku yimid hidda-wadaha 423-ka ee MLP-ka ayaa la arkay inta lagu jiro difaaca avokado ee caabuqa F. niger iyo inta lagu jiro caabuqa geedka tufaaxa Botryosphaeria berengeriana f. cn. piricola iyo Alternaria alternata waa noocyada tufaaxa58,59. Intaa waxaa dheer, calli-ka tufaaxa ee si xad dhaaf ah u muujiya hidda-wadaha 423-ka ee MLP-ka ayaa lahaa muujinta hoose ee hidda-wadaha la xiriira iska caabbinta waxayna u nugul yihiin caabuqa fungal59. Ka dib Fusarium oxysporum f, hidda-wadaha 423-ka ee MLP-ka ayaa sidoo kale la xakameeyay jeermiska digirta caadiga ah ee u adkaysta. cn. Cudurka digirta 60.
Xubnaha kale ee qoyska PR-10 ee lagu aqoonsaday daraasaddeenna RNA-seq waxay ahaayeen hiddo-sideyaasha LlR18A iyo LlR18B ee ka jawaabaya kor u qaadista, iyo sidoo kale hiddo-sideyaasha kor u kacay (1 hiddo-side) ama kuwa hoos u dhacay (3 hiddo-side) ee borotiinka wareejinta dufanka DIR1. . Intaa waxaa dheer, WGCNA waxay iftiiminaysaa hiddo-sideyaasha LlR18B inay yihiin xarun ku jirta qaybtan, kaas oo aad ugu nugul tallaalka isla markaana wata dhowr hiddo-sideyaal jawaab celin ilaalin ah. Hiddo-sideyaasha LlR18A iyo LlR18B waxaa lagu kiciyay caleenta lupine ee jaalaha ah iyagoo ka jawaabaya bakteeriyada cudurada keena, iyo sidoo kale jirridda NLL ka dib tallaalka D. toxica, halka isku-dhafka bariiska ee hiddo-sideyaashan, RSOsPR10, si degdeg ah uu u kiciyay caabuq fangas ah oo loo maleynayo inuu ku lug lahaa waddada calaamadaynta aashitada jasmonic53,61, 62. Hiddo-sideyaasha DIR1 waxay codeeyaan borotiinnada gaadiidka lipid-ka ee aan gaarka ahayn ee loo baahan yahay bilowga iska caabinta nidaamsan (SAR). Iyadoo la horumarinayo falcelinta ilaalinta, borotiinka DIR1 waxaa laga qaadaa diiradda caabuqa iyada oo loo marayo phloem si loogu kiciyo SAR xubnaha fog. Waxaa xiiso leh, hidda-wadaha TanjilG_02313 DIR1 ayaa si weyn loo kiciyay markii ugu horreysay ee xariiqyada 84A:476 iyo Dadweynaha 22660, laakiin iska caabbinta anthracnose waxay si guul leh ugu soo baxday oo keliya xariiqda 84A:476. Tani waxay muujin kartaa waxoogaa hoos u dhac ah oo ku yimid hidda-wadaha DIR1 ee NLL, maadaama saddexda isku-dhafan ee soo haray ay ka jawaabeen tallaalka oo keliya xariiqda 83A:476 ee 6 hpi, jawaabtanna waxaa loo jeediyay hoos.
Daraasaddeenna, qaybaha ugu badan ee u dhigma habka bayoolojiga ee loo yaqaan "GO:0055114 Redox process" waxay ahaayeen borotiinka cytochrome P450, peroxidase, linoleic acid 9S-/13S-lipoxygenase, iyo 1-aminocyclopropane-1-carboxylic acid oxidase. Intaa waxaa dheer, WGCNA-gayaga wuxuu qeexayaa isku-dhafka HSFA4a inuu yahay qaybo sita xarun sida musharaxa hidda-wadaha iska caabbinta Lanr1 TanjilG_05042. HSFA4a waa qayb ka mid ah nidaaminta ku-tiirsanaanta redox ee qoraalka nukliyeerka ee dhirta.
Borotiinnada Cytochrome P450 waa oksaydh-oxidoreductases kuwaas oo kiciya falcelinta hydroxylation ee NADPH iyo/ama O2-ku-tiirsan ee dheef-shiid kiimikaadka aasaasiga ah iyo kan sare, oo ay ku jiraan dheef-shiid kiimikaadka xenobiotics, iyo sidoo kale hoormoonnada, asiidhyada dufanka leh, sterols, qaybaha derbiga unugyada, biopolymers, iyo bayoolojiyada isku-dhafka ilaalinta 69. Daraasaddeenna, kala duwanaanshaha shaqada cytochrome P450 ee dhirta ayaa laga dhimay -10.6 log2 (isbeddel laab ah) ilaa 5.7 sababtoo ah tiro badan oo isku-dhafan oo isbeddelay (37) iyo kala duwanaanshaha qaababka jawaab celinta ee u dhexeeya hiddo-wadaha gaarka ah, oo ka tarjumaya dib-u-eegis kor u kacay. . Isticmaalka xogta RNA-seq oo keliya si loo caddeeyo shaqada bayoolojiga ee hidde-sideyaasha NLL ee qoyska sare ee borotiinka ee noocaas ah waxay noqon doontaa mid aad u male-awaal ah. Si kastaba ha ahaatee, waxaa xusid mudan in qaar ka mid ah hiddo-wadaha cytochrome P450 ay la xiriiraan iska caabinta sii kordheysa ee fangaska ama bakteeriyada cudurada keena, oo ay ku jiraan gacan ka geysashada falcelinta xasaasiyadda69,70,71.
Fasalka III peroxidases waa ensaymo dhir oo badan oo ku lug leh noocyo badan oo geedi socod dheef-shiid kiimikaad ah inta lagu jiro koritaanka iyo horumarka dhirta, iyo sidoo kale jawaab celinta walbahaarka deegaanka sida milixda, abaarta, xoogga iftiinka sare, iyo weerarka jeermiska72. Peroxidases waxay ku lug leeyihiin isdhexgalka noocyo badan oo dhir ah oo la leh Anthracis, oo ay ku jiraan Stylosanthes humilis iyo C. gloeosporioides, Lens culinaris iyo C. truncatum, Phaseolus vulgaris iyo C. lindemuthianum, Cucumis sativus iyo C. lagenarium73,74,75,76. Jawaabtu aad bay u degdeg badan tahay, mararka qaarkood xitaa 4 HPI, ka hor inta uusan fangaska gelin unugyada dhirta73. Hidde-sidaha peroxidase sidoo kale wuxuu ka jawaabay tallaalka D. toxica NLL. Marka lagu daro shaqooyinkooda caadiga ah si ay u xakameeyaan qarxinta oksaydhka ama ay u baabi'iyaan walbahaarka oksaydhka, peroxidases waxay faragelin karaan koritaanka jeermiska iyagoo abuuraya caqabado jireed oo ku salaysan xoojinta derbiga unugyada inta lagu jiro isku-xidhka, qaybta hoose ama isku-xirka isku-xidhka xeryahooda gaarka ah. Shaqadan waxaa lagu tirin karaa silico hidda-wadaha TanjilG_03329 ee codeeya anion peroxidase-ka sameeya lignin-ka kaasoo si weyn kor loogu qaaday daraasaddeenna ku jirta xariiqda u adkaysata 83A:476 ee 6 HPI, laakiin aan ku jirin noocyada kale iyo waqtiyada aan ka jawaabin.
9S-/13S-lipoxygenase ee linoleic acid waa tallaabada ugu horreysa ee marinnada oksaydhka ee biosynthesis-ka lipid78. Waxyaabaha ka soo baxa marinnadan waxay leeyihiin shaqooyin badan oo difaaca dhirta ah, oo ay ku jiraan xoojinta derbiga unugyada iyada oo loo marayo sameynta kaydadka callose iyo pectin, iyo nidaaminta walbahaarka oksaydhka iyada oo loo marayo soo saarista noocyada oksijiinta ee falcelinta leh79,80,81,82,83. Daraasaddan hadda socota, muujinta aashitada linoleic 9S-/13S-lipoxygenase ayaa wax laga beddelay dhammaan noocyada, laakiin dadka nugul 22660, kor u kaca ayaa ka sarreeyay waqtiyo kala duwan, halka noocyada sita Lanr1 u adkaysta iyo allele AnMan, ay xoogga saarayso kala duwanaanshaha lakabka oxylipin ee falcelinta anthrax ee ilaalinta u dhexeeya noocyadan.
Is-dhexgalka 1-aminocyclopropane-1-carboxylate oxidase (ACO) ayaa si weyn kor loogu qaaday (9 hidde) ama hoos loo dhigay (2 hidde) markii lagu tallaalay lupine. Laba ka reeban, dhammaan jawaabahan waxay dhaceen 6 hp. 83A:476. Falcelinta ensaymikada ee ay dhexdhexaadisay borotiinnada ACO waa tallaabada xaddidaysa heerka wax soo saarka ethylene sidaas darteedna si aad ah ayaa loo nidaamiyay84. Ethylene waa hoormoon dhir ah oo ka ciyaara doorar kala duwan oo ku saabsan nidaaminta horumarinta dhirta iyo ka jawaabista xaaladaha walaaca abiotic iyo biotic. Soo saarista qoraalka ACO iyo kicinta wadada calaamadaynta ethylene waxay ku lug leedahay kordhinta iska caabbinta bariiska ee fangaska hemibiotrophic oryzae oryzae iyadoo la nidaaminayo soo saarista noocyada oksijiinta ee falcelinta leh iyo phytoalexins. Geedi socod aad u la mid ah oo caabuqa caleenta ah oo laga helay inta u dhaxaysa M. oryzae iyo C. lupini88,89, iyadoo laga soo horjeedo kor u kaca weyn ee isku-dhafka ACO ee xariiqda 83A:476 ee lagu soo sheegay daraasaddan, waxay beddeshaa suurtagalnimada in la siiyo iska caabbinta anthracnose NLL Ethylene tallaabo dhexe oo calaamad u ah waddooyinka molecular-ka.
Daraasaddan hadda socota, caburin ballaaran oo hidde-sideyaal badan oo la xiriira sawir-qaadista ayaa lagu arkay 6 hpi 83A:476 iyo 48 hpi Mandeloop iyo 22660 dadweynaha. Baaxadda iyo horumarka isbeddelladani waxay u dhigantaa heerka. Iska caabbinta Anthracnose ayaa lagu arkay tijaabadan. Dhawaan, caburin xooggan oo hore oo ku saabsan qoraallada la xiriira sawir-qaadista ayaa lagu soo warramey dhowr nooc oo isdhexgalka dhirta iyo cudur-sidaha, oo ay ku jiraan bakteeriyada cudurka keena iyo fangaska. Degdegga (laga bilaabo 2 HPI isdhexgalka qaar) iyo caburinta caalamiga ah ee hidde-sideyaasha la xiriira sawir-qaadista si looga jawaabo caabuqa waxay kicin kartaa difaaca dhirta iyadoo lagu saleynayo dejinta noocyada oksijiinta ee falcelinta leh iyo isdhexgalka ay la leeyihiin marinka salicylic acid si loo dhexdhexaadiyo falcelinta xasaasiyadda 90,94.
Gunaanad ahaan, hababka jawaab celinta difaaca ee loo soo jeediyay nooca ugu adkeysiga badan (83A:476) waxaa ka mid ah aqoonsashada degdegga ah ee cudur-sidaha ee hidda-wadaha R (malaha TIR-NBS-LRR TanjilG_05042) iyo salicylic acid iyo ethylene oo ay dhexdhexaadiso jawaab celinta xasaasiyadda, oo ay ku xigto sameynta ficilka SAR ee fog. Waxaa taageera borotiinka DIR-1. Waa in la ogaadaa in muddada bayotrophic ee caabuqa C. lupini uu aad u gaaban yahay (qiyaastii 2 maalmood), oo ay ku xigto koritaanka necrotic95. Kala-guurka u dhexeeya marxaladahaan waxaa lala xiriirin karaa necrosis iyo muujinta borotiinnada ethylene-ka ee la kicin karo kuwaas oo u dhaqma sidii kiciyayaal falcelin xasaasiyadeed oo ku dhaca dhirta martida loo yahay. Sidaa darteed, waqtiga daaqadda loogu talagalay qabashada guusha leh ee C. lupini marxaladda bayotrophic waa mid aad u cidhiidhi ah. Dib-u-habaynta hidda-wadaha la xiriira redox iyo photosynthesis ee lagu arkay 83A:476 6 hpi waxay la jaanqaadaysaa horumarka hyphae-ka fangaska waxayna ku dhawaaqdaa horumarinta jawaab celin difaac oo guul leh marxaladda bayotrophic. Jawaabaha qoraalka ah ee Mandelup iyo dadka 22660 ayaa laga yaabaa inay aad u daahdo si ay u qabtaan fangaska ka hor inta aysan u wareegin koritaanka necrotic, si kastaba ha ahaatee, Mandelup waxay ka waxtar badan tahay dadka 22660 sababtoo ah nidaaminta degdegga ah ee borotiinka PR-10 waxay kor u qaadaysaa iska caabinta toosan.
ETI, oo ay wado hidda-wadaha R ee caadiga ah, waxay u muuqataa inay tahay farsamo caadi ah oo loogu talagalay iska caabbinta digirta ee anthracnose. Sidaa darteed, qaabka geedka Medicago truncatula, iska caabbinta anthracnose waxaa bixiya hidda-wadaha RCT1, oo xubin ka ah fasalka hidda-wadaha R ee dhirta TIR-NBS-LRR97. Hidda-wadahani waxay sidoo kale bixisaa iska caabbinta anthracnose ee ballaaran ee alfalfa marka loo wareejiyo dhirta nugul. Digirta caadiga ah (P. vulgaris), in ka badan laba iyo toban hiddo-wade oo iska caabbinta anthracnose ayaa ilaa hadda la aqoonsaday. Qaar ka mid ah hidda-wadahani waxaa laga helaa gobollada aan lahayn hidda-wade R oo caan ah, si kastaba ha ahaatee kuwo kale oo badan ayaa ku yaal cidhifyada koromosoomyada sita kooxda hidda-wadaha NBS-LRR, oo ay ku jiraan TIR-NBS-LRRs99. Daraasadda SSR ee genome-ka oo dhan ayaa sidoo kale xaqiijisay xiriirka hidda-wadaha NBS-LRR iyo iska caabbinta anthracnose ee digirta caadiga ah. Hidde-sidayaasha R ee canonical-ka ah ayaa sidoo kale laga helay gobolka genomic-ga oo sita meesha ugu weyn ee iska caabbinta anthracnose ee ku taal lupine cad 101.
Shaqadeennu waxay muujinaysaa in falcelin iska caabin degdeg ah, oo la hawlgeliyo marxalad hore ee caabuqa dhirta (ugu yaraan 12 hpi), ay si wax ku ool ah uga ilaaliso lupine-ka caleenta cidhiidhiga ah anthracnose oo uu keeno fangaska bakteeriyada Collelotrichum lupini. Annagoo adeegsanayna isku xigxiga sare ee wax soo saarka, waxaan muujinnay muuqaallo kala duwan oo muujinaya hiddo-sideyaasha iska caabbinta anthracnose ee dhirta NLL oo ay dhexdhexaadiyeen hiddo-sideyaasha iska caabbinta Lanr1 iyo AnMan. Difaaca guuleysta wuxuu ku lug leeyahay si taxaddar leh u qaabaynta hiddo-sideyaasha borotiinnada ku lug leh redox, photosynthesis, iyo pathogenesis-ka saacado gudahood marka geedka uu la xiriiro cudur-sidaha. Falgallada ilaalinta ee la midka ah, laakiin dib u dhaca waqtiga, aad bay uga waxtar yar yihiin ilaalinta dhirta cudurrada. Iska caabbinta Anthrax oo ay dhexdhexaadiso hiddo-sideyaasha Lanr1 waxay u egtahay jawaabta degdegga ah ee caadiga ah ee hiddo-sideyaasha R (difaaca kiciya saamaynta), halka hiddo-sideyaasha AnMan ay u badan tahay inay bixiyaan jawaab toosan (difaaca oo ay kiciyaan qaab molecular ah oo microbe-la xiriira), iyagoo bixinaya heer dhexdhexaad ah oo waara.
215-ka xariiq ee NLL ee loo isticmaalay in lagu baaro calaamadaha anthracnose waxay ka koobnaayeen 74 nooc, 60 xariiq oo laga helay isgoys ama taranta, 5 isbeddel hidde-sideyaal ah, iyo 76 jeermis oo duurjoog ah ama asal ah. Khadadka waxay ka yimaadeen 17 waddan, oo inta badan ka yimid Poland (58), Spain (47), Jarmalka (27), Australia (26), Ruushka (19), Belarus (7), Talyaaniga (5) iyo xariiqyo kale. Qaybtani waxay sidoo kale ka kooban tahay xariiqyo u adkaysta tixraaca: 83A:476, Tanjil, Wonga oo sidda allele Lanr1, iyo Mandelup oo sidda allele AnMan. Khadadka waxaa laga helay Kaydka Kheyraadka Hidde-sidaha ee Yurub ee Lupine oo ay ilaaliso Poznań Plant Breeding Ltd., Wiatrowo, Poland (Jadwalka Dheeraadka ah S1).
Dhirta waxaa lagu beeray xaalado la xakameeyey (fotoperiode 16 saacadood, heerkulka 25°C maalintii iyo 18°C ​​habeenkii). Laba nuqul oo bayooloji ah ayaa la falanqeeyay. DNA-da waxaa laga soocay caleemo saddex toddobaad jir ah iyadoo la adeegsanayo Qalabka Yar ee DNeasy Plant (Qiagen, Hilden, Jarmalka) sida ku cad hab-maamuuska. Tayada iyo xoojinta DNA-da go'doonsan waxaa lagu qiimeeyay hababka spectrophotometric (NanoDrop 2000; Thermo Fisher Scientific, Waltham, MA, USA). Calaamadda AnManM1 ee calaamadaynaysa hidda-wadaha iska caabbinta anthracnose AnMan (oo laga soo qaatay cv. Mandelup) iyo calaamadaha Anseq3 iyo Anseq4 ee ku teedsan hidda-wadaha Lanr1 (oo laga soo qaatay cv. Tanjil) ayaa la falanqeeyay 11,26,28. Homozygotes-ka allele-ka iska caabbinta waxaa lagu qiimeeyay "1″, oo u nugul - "0″, iyo heterozygotes - oo ah 0.5.
Iyada oo lagu saleynayo natiijooyinka baaritaanka calaamadaha AnManM1, AnSeq3 iyo AnSeq4 iyo helitaanka abuurka tijaabooyinka dabagalka ugu dambeeya, 50 xariiq oo NLL ah ayaa loo xushay phenotyping-ka iska caabbinta anthracnose. Falanqaynta waxaa lagu sameeyay nuqul ka mid ah aqalka dhirta lagu koriyo oo kombiyuutarku maamulo oo leh muddo 14 saacadood ah oo sawir-qaadis ah oo leh heerkul ah 22°C maalintii iyo 19°C habeenkii. Abuurka waa la xoqaa (laga jaraa daboolka abuurka dhinaca ka soo horjeeda ee uurjiifka iyadoo la isticmaalayo daab fiiqan) ka hor inta aan la beerin si looga hortago abuurka inuu seexdo sababtoo ah dahaarka abuurka oo aad u adag iyo in la hubiyo biqilka isku mid ah. Dhirta waxaa lagu beeray dheriyo (11 × 11 × 21 cm) oo leh ciid nadiif ah (TS-1 REC 085 Medium Basic, Klasmann-Deilmann Polska, Warsaw, Poland). Tallaalka waxaa lagu sameeyay nooca Colletotrichum lupini Col-08, kaas oo la beeray 1999 laga bilaabo jirridaha dhirta lupine ee caleenta cidhiidhiga ah ee lagu beeray beer ku taal Verzhenitsa, Greater Poland (52° 27′ 42″ N 17° 04′ 05″ E). Hel meel. Go'doominta waxaa lagu beeray meel dhexdhexaad ah oo SNA ah 20° C. iyadoo la adeegsanayo iftiin madow muddo 21 maalmood ah si loo kiciyo sporulation. Afar toddobaad ka dib beerista, marka dhirtu gaarto marxaladda caleen 4-6, tallaalka waxaa lagu sameeyay iyadoo lagu buufinayo joojinta conidia oo ah 0.5 x 106 conidia halkii ml. Ka dib markii la tallaalay, dhirta waxaa lagu hayay mugdi 24 saacadood iyadoo qoyaankeedu yahay qiyaastii 98% iyo heerkul ah 25°C si loo fududeeyo biqilka conidia iyo habka caabuqa. Dhirta waxaa lagu beeray muddo 14 saacadood ah oo sawir-qaadis ah habeenkii 22°C/19°C iyo 70% qoyaan. Dhibcaha cudurka waxaa la sameeyay 22 maalmood ka dib tallaalka waxayna u dhaxaysay 0 (difaac) ilaa 9 (aad u nugul) iyadoo ku xiran jiritaanka ama maqnaanshaha nabarrada necrotic-ga ee jirridda iyo caleemaha. Intaa waxaa dheer, ka dib markii la qiimeeyay, miisaanka dhirta ayaa la cabbiray. Xiriirka ka dhexeeya noocyada calaamadaynta iyo noocyada cudurka waxaa lagu xisaabiyay isku xidhka laba-taxane ee dhibicda (maqnaanshaha calaamadaha heterozygous ee xariiqyada loogu talagalay falanqaynta nooca iska caabbinta anthracnose).