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Angustifolius lupine (NLL, Lupinus angustifolius L.) waa geed leguminous ah oo loo isticmaalo wax soo saarka cuntada iyo hagaajinta ciidda.Balaadhinta caalamiga ah ee NLL sida dalagga ayaa soo jiidatay fangaska badan ee cudur-sidaha, oo ay ku jiraan lupine anthracnose, kaas oo sababa cudurka halista ah ee anthracnose.Laba alleles, Lanr1 iyo AnMan, kuwaas oo keena caabbinta korodhay, ayaa loo adeegsaday taranka NLL, laakiin hababka unugyo ee hoose ayaan la garanayn.Daraasaddan, calaamadaha Lanr1 iyo AnMan ayaa loo isticmaalay in lagu baaro muunadaha NLL ee Yurub.Tijaabada tallaalka ee deegaan kontorool lagu sameeyay ayaa xaqiijisay waxtarka deeq bixiyayaasha iska caabinta ah.Soo bandhigida muujinta hidde-sidaha kala duwan ayaa lagu sameeyay xariiqyo adkaysi u leh oo u nugul wakiillada.Caabbinta Anthracnose waxay la xiriirtay si xad dhaaf ah oo ku saabsan ereyada hidda-socodka ontology "GO: 0006952 Jawaabta Difaaca", "GO: 0055114 Nidaamka Dib-u-habeynta", iyo "GO: 0015979 Photosynthesis".Intaa waxaa dheer, khadka Lanr1 (83A: 476) wuxuu muujiyay dib u habeyn weyn oo qoraal ah oo degdeg ah ka dib tallaalka, halka khadadka kale ay muujiyeen dib u dhac ku yimid jawaabtan ilaa 42 saacadood.Jawaabaha difaaca waxay la xiriiraan TIR-NBS, CC-NBS-LRR iyo hiddo-wadaha NBS-LRR, borotiinnada 10 ee ku lug leh pathogenesis, borotiinka wareejinta dufanka, endoglucan-1,3-β-glucosidase, borotiinnada gidaarka unugyada glycine-hodan, iyo hiddo-wadaha ka soo jeeda dariiqa falcelinta ee oksijiinta.Jawaabaha hore ee 83A: 476, oo ay ku jiraan xakamaynta taxaddarka leh ee hiddo-wadaha la xidhiidha photosynthesis, waxay ku beegan tahay ilaalinta guusha leh inta lagu jiro marxaladda koritaanka daaqsinka ee bayoolajiga fangas, taas oo soo jeedinaysa in saameyn leh uu kiciyo difaac.Dareen-celinta Mandeloop waa la dhimay, sida guud ahaan jiidashada toosan.
Lupine-caleemo cidhiidhi ah (NLL, Lupinus angustifolius L.) waa badarka borotiinka sare leh oo ka soo jeeda galbeedka Mediterranean-ka gobolka1,2.Hadda waxa loo beeraa dalagga cunto ee xoolaha iyo dadka.Waxa kale oo loo arkaa digada cagaaran ee hababka wareegga dalagga ay sabab u tahay hagaajinta nitrogen by symbiotic nitrogen hagaajinta bakteeriyada iyo horumarinta guud ee dhismaha ciidda.NLL waxa ay soo martay hannaan deg-deg ah oo guri-gurid qarnigii la soo dhaafay waxana uu weli ku jiraa cadaadis sare oo taranka3,4,5,6,7,8,9,10,11,12.Iyada oo beerashada baahsan ee NLL, isku xigxiga ee fangaska cudur-sidaha waxay horumariyeen beero beero cusub waxayna sababeen cudurro cusub oo baabi'iya dalagga. Midda ugu cajiibsan ee beeralayda lupine iyo taranta waxay ahayd muuqaalka kudka, oo ay sababto fungus pathogenic, Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Midda ugu cajiibsan ee beeralayda lupine iyo taranta waxay ahayd muuqaalka kudka, oo ay sababto fungus pathogenic, Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Наиболее примечательным для фермеров и селекционеров люпина lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Waxa ugu caansan beeralayda iyo taranayaasha lupine waxa ay ahayd soo ifbaxa anthracnose oo uu sababay fangaska dilaaga ah 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，它是由病原真語由病原真说！ 3 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 totrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Inta ugu badan ee beeralayda lupine iyo taranta waa soo bixitaanka anthracnose ee uu keeno fungus pathogenic Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13.Warbixinadii ugu horreeyay ee cudurkan ayaa ka yimid Brazil iyo Mareykanka, iyadoo calaamadaha caadiga ah ay soo baxeen 1912 iyo 1929 siday u kala horreeyaan.Si kastaba ha ahaatee, qiyaastii 30 sano ka dib, cudur-sidaha waxaa loo qoondeeyey Colletotrichum gloeosporioids (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 in Morphology Bartilmaameedka. & H. Schrenk,. & H. Schrenk, .iyo H. Schrenk. & H.施伦克，. & H.施伦克，.iyo H. Schlenk,.Cudurka fenotyping ee horudhaca ah ee la sameeyay bartamihii qarnigii 20aad ayaa muujiyay xoogaa iska caabin ah NLL iyo lupine jaalaha ah (L. luteus L.) gelitaanka, laakiin dhammaan noocyada 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 kordhay (huurka hawada) iyo heerkulka (ee kala duwan ee 12-28 ° C), taasoo keentay xadgudub ku ah caabbinta heerkulka sare17, 18. Dhab ahaantii, waqtiga loo baahan yahay conidia biqilka iyo cudurka si uu u bilaabo , ahaa afar jeer ka gaaban at 24 ° C (4 saacadood) ka hooseeya 1 xaaladaha (4 saacadood) 1.Sidaa darteed, kulaylka caalamiga ah ee socda ayaa horseeday fidinta anthracnose.Si kastaba ha ahaatee, cudurkan waxaa lagu arkay Faransiiska (1982) iyo Ukraine (1983) oo ah khatar khatar ah oo soo socota, laakiin sida muuqata waxaa iska indhatiray warshadaha lupine waqtiga20,21.Dhowr sano ka dib, cudurkan halista ah ayaa ku fiday adduunka oo dhan wuxuuna sidoo kale saameeyay waddamo waaweyn oo soo saara Lupine sida Australia, Poland iyo Germany22,23,24.Ka dib markii uu dillaacay anthracnose badhtamihii 1990-aadkii, baadhis ballaadhan ayaa keentay in la ogaado deeq-bixiyeyaal badan oo adkaysi u leh muunado NLL19 ah.NLL caabbinta anthracnose waxaa gacanta ku haya laba alleles oo kala duwan oo laga helo ilo jeermisyo kala duwan: Lanr1 oo ku yaal cultivar Tanjil iyo Wonga iyo AnMan ee cultivar.Mandalay 25, 26. alleles-yadani waxay dhamaystirayaan calaamadaha molecular ee taageera doorashada jeermiska u adkaysta ee barnaamijyada taranka25,26,27,28,29,30.Xariiqda taranka ee adkaysiga leh ee 83A: 476 ee sidda Lanr1 allele ayaa laga gudbay khadka duurjoogta ah ee nugul P27255 si loo helo tirada dadka RIL ee u kala soocida iska caabbinta anthracnose, taas oo suurtogal ka dhigtay in lagu meeleeyo goobta Lanr1 koromosoom NLL-1131, 32, 33 NLL ayaa shaaca ka qaaday meesha ay ku yaalliin dhammaan saddexda alleles ee ku jira koromosoomyada isku midka ah (NLL-11), laakiin ku yaal meelo kala duwan29,34,35.Si kastaba ha ahaatee, iyadoo ay ugu wacan tahay tirada yar ee RIL-yada iyo masaafada weyn ee hidde-sidaha u dhexeeya calaamadaha iyo alleles u dhigma, ma jirto gabagabo la isku halayn karo oo laga soo saari karo hiddo-wadaha hoose.Dhanka kale, isticmaalka hidda-socod-celinta ee lupins-ka way adagtahay sababtoo ah awooddooda dib-u-soo-nooleynta oo aad u hooseeya, taas oo ka dhigaysa khalkhalgelinta hidde-sidaha37.
Kobcinta jeermiska guriga ee sidda allele la rabo ee gobolka qaniisyada, sida 83A: 476 (Lanr1) iyo Mandelup (AnMan), ayaa albaabka u furay in la barto iska caabbinta anthracnose iyada oo ay jirto joogitaanka isku-dhafka alleles ee dadka duurjoogta ah.Fursadaha hababka molecular.Is barbar dhig jawaabaha difaaca ee ay keeneen genotypes gaar ah.Daraasadani waxay qiimeysay jawaabta hore ee qoraalka ee NLL ilaa C. lupini tallaalka.Marka hore, guddi jeermis ah oo NLL ah oo Yurub ah oo ka kooban 215 xariiq ayaa la baadhay iyadoo la isticmaalayo calamadaha molecular ee calaamadeynaya Lanr1 iyo AnMan alleles.Anthracnose phenotyping ayaa markaa lagu sameeyay khadadka 50 NLL, oo ​​hore loogu doortay calaamadaha molecular, ee xaaladaha la xakameeyey.Iyada oo ku saleysan tijaabooyinkan, afar xariiq oo ku kala duwan iska caabbinta anthracnose iyo Lanr1/AnMan allelic ayaa loo xushay muujinta hidde-sidaha difaaca kala duwan iyadoo la adeegsanayo laba hab oo dhammaystiran: isku xigxiga RNA-sare oo la soo saaray iyo cabbir PCR-waqtiga-dhabta ah.
Baaritaanka jeermiska NLL ee NLL (N = 215) oo leh calaamadeeyayaal Lanr1 (Anseq3 iyo Anseq4) iyo AnMan (Anseq4) iyo AnMan (AnManM1) waxay muujisay in hal xariiq oo keliya (95726, u dhow Salamanca-b) ay kor u qaadayso "iska caabinta" allele ee dhammaan calaamadaha. 73.5%) khadadka.Saddex iyo toban xariiq ayaa soo saaray laba alleles oo "iska caabiya" calaamada Lanr1, 8 xariiquna waxa ay soo saareen alleles "iska adkaysta" ee Lanr1.calaamadeeye.Calaamadaha "iska caabinta" ee calaamadeeyaha AnMan (Shaxda Dheeraadka ah S1).Laba sadar waxay ahaayeen heterozygous calaamadeeyaha Anseq3 iyo hal heterozygous ee calaamadeeyaha AnManM1.42 xariiq (19.5%) ayaa qaaday wejiyaal iska soo horjeeda oo ah Anseq3 iyo Anseq4 alleles, taas oo muujinaysa soo noqnoqoshada badan ee dib-u-habaynta inta u dhaxaysa labadan goobood.Noocyada Anthracnose ee xaaladaha la xakameeyey (Shaxda Dheeraadka ah S2) ayaa muujiyay kala duwanaanshiyaha caabbinta genotypes-ka la tijaabiyay, taas oo ka muuqatay darnaanta anthracnose.Farqiga u dhexeeya dhibcaha celceliska ayaa u dhexeeyay 1.8 (dhexdhexaad u adkaysta) ilaa 6.9 (u nugul) iyo kala duwanaanshaha miisaanka dhirta ayaa u dhexeeyay 0.62 (u nugul) ilaa 4.45 g (u adkaysta). Waxaa jiray xiriir weyn oo u dhexeeya qiyamka lagu arkay laba nooc oo tijaabo ah (0.51 ee buundooyinka darnaanta cudurka, P = 0.00017 iyo 0.61 ee miisaanka dhirta, P <0.0001) iyo sidoo kale inta u dhaxaysa labadan cabbir (- 0.59 iyo - 0.77, P <0.0001). Waxaa jiray xiriir weyn oo u dhexeeya qiyamka lagu arkay laba nooc oo tijaabo ah (0.51 ee buundooyinka darnaanta cudurka, P = 0.00017 iyo 0.61 ee miisaanka dhirta, P <0.0001) iyo sidoo kale inta u dhaxaysa labadan cabbir (- 0.59 iyo - 0.77, P <0.0001). 0,51 . 0,00017 iyo 0,61 для массы растения, P <0,0001), а также межdu эtymy дvyumya parametram (-0,59 и -0,77, 0,00) 0,0). Xiriir weyn ayaa laga helay inta u dhaxaysa qiyamka lagu arkay laba jeer oo tijaabo ah (0.51 ee buundooyinka darnaanta cudurka, P = 0.00017 iyo 0.61 ee miisaanka dhirta, P <0.0001), iyo sidoo kale inta u dhaxaysa labadan cabbir (- 0.59 iyo -0.77, P <0.0001) 1).. 0.61, P <0.0001）以及这两个参数之间（- 0.59 和- 0.77，P <0.0001)在 两 次重复 实验 中 观察 的 值 之间 存在 相关性0.00017 ， 植物 为 为 0.61和 - 0.77, P <0.0001). Наблюдалась значительная корреляция между значениями 017 iyo mасса растения 0,61, P <0,0001), iyo межdu эtymy dvyumya parametram (-0,59 и -0,0001) 0,77, P <0,0001. Waxaa jiray xiriir weyn oo u dhexeeya qiyamka lagu arkay nuqul ka mid ah (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 jilbiska iyo qalloocinta jirridda oo u eg qaab-dhismeedka " qaansada adhijirka ", oo ay ku xigto nabarro oval ah oo leh sporozoites orange / casaan (Jaantus Dheeraad ah 1).Galitaanka Australiyaanka ee sida Lanr1 (83A: 476 iyo Tanjil) iyo AnMan (Mandelup) hiddo-wadaha ayaa si dhexdhexaad ah u adkaysta, 0.0331 iyo 0.0036).Qaar ka mid ah khadadka kuwaas oo sidoo kale sita "adkeysi" Lanr1 iyo/ama AnMan alleles ayaa muujinaya calaamadaha cudurka.
Waxa xiiso leh, dhowr xariiq oo NLL ah oo ka maqan calaamad kasta oo "adkeysi leh" ayaa muujisay heer sare oo caabbinta anthracnose ah (la barbardhigi kara ama ka sarreeya Lanr1 ama AnMan genotypes), sida Boregine (qiimaha P <0.0001 labadaba), Bojar (qiimaha P <0.0001 ee dhibcaha iyo 0.0001 ee miisaanka 0.001 ee miisaanka B000) oo aan miisaan ku lahayn). Waxa xiiso leh, dhowr xariiq oo NLL ah oo ka maqan calaamad kasta oo "adkeysi leh" ayaa muujisay heer sare oo caabbinta anthracnose ah (la barbardhigi kara ama ka sarreeya Lanr1 ama AnMan genotypes), sida Boregine (qiimaha P <0.0001 labadaba), Bojar (qiimaha P <0.0001 ee dhibcaha iyo 0.0001 ee miisaanka 0.001 ee miisaanka B000) oo aan miisaan ku lahayn). Интересно, что несколько линий NLL, лишенных какого-либо ости к антракнозу ), Bojar (значение P <0,0001 для оценки и 0,001 ). Waxa xiiso leh, dhowr xariiq oo NLL ah oo ka maqan calaamad kasta oo 'adkeysi leh' ayaa muujisay heer sare oo iska caabin ah oo ku saabsan anthracnose (marka la barbardhigo ama ka sarreeya Lanr1 ama AnMan genotypes), sida Boregine (qiimaha P <0.0001 labadaba), Bojar (qiimaha P <0.0001 ee qiimaynta iyo 0.0001 dhirta) 1 si loo qiimeeyo oo aan muhiimad u lahayn miisaanka).有趣的是，一些缺乏任何“抗性”标记等位基因的NLL型相当或更高），例如Boregine（两个参数的P 值< 0.0001）、Bojar（P 值<得分丏为0.0001，得分丏0.0001，得分丏0.0001） B-549/79b（得分P 值< 0.0001，重量不显着）。 Waxaa xiiso leh in qaar ka mid ah nidaamyada NLL ee aan haysan wax calaamado "antigenic" ah ay muujinayaan iska caabin sare oo toosan (oo u dhiganta Lanr1 ama hiddo-wadaha AnMan ama ka sareeya), sida Boregine (labada cabbir ee P <0.0001), Bojar (qiimaha P <0.0001, miisaanka dhirta 0.001) iyo cadaadiska B-549 / 0.0.0. Интересно, что некоторые линии NLL. к антракнозу (сравнимые или выше, чем у генотипов Lanr1 или AnMan), такие как Boregine 1, масса растения 0,001) iyo популяция B-549/79b (оценка P-значение <0,0001, масса незначительна). Waxa xiisaha lihi leh, qaar ka mid ah khadadka NLL ee aan lahayn wax kasta oo 'caabbinta' ah ayaa muujinaya heerar sare oo caabbinta anthracnose ah (marka la barbar dhigo ama ka sarreeya Lanr1 ama genotypes AnMan), sida Boregine (P qiimaha labada cabbir <0.0001) , Bojar (P-qiimaha <0.0001, miisaanka dhirta 0.09-051) miisaan aan muhiim ahayn).Dhacdadani waxay soo jeedinaysaa suurtagalnimada ilo cusub oo hidde-celin ah oo iska caabin ah, taasoo sharraxaysa xiriir la'aanta la arkay ee u dhexeeya calaamadaha genotypes iyo phenotypes cudurada (qiimaha P ee ~ 0.42 ilaa ~ 0.98).Sidaa daraadeed, imtixaanka Kolmogorov-Smirnov wuxuu muujiyay in xogta ku saabsan iska caabbinta anthracnose lagu qiyaaso sida caadiga ah loo qaybiyo dhibcaha (P-qiimaha 0.25 iyo 0.11) iyo xajmiga dhirta (P-qiimaha 0.47 iyo 0.55), oo soo jeedinaya in aan qiyaaso in ka badan oo ka badan Lanr1 iyo AnMan ay ku lug leeyihiin.
Iyada oo ku saleysan natiijooyinka baaritaanka iska caabbinta anthracnose, 4 xariiq ayaa loo xushay falanqaynta transcriptome: 83A: 476, Boregine, Mandelup, iyo Population 22660. Khadadkan ayaa dib loo tijaabiyay caabbinta kudka ee tijaabooyinka tallaalka ee isku xigxiga RNA, waase haddii ay la mid yihiin imtixaankii hore.Qiimaha dhibcaha waxay ahaayeen sida 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).
Nidaamka Illumina NovaSeq 6000 wuxuu gaadhay celcelis ahaan 40.5 Mread lammaane muunad kasta (29.7 ilaa 54.4 Mreads) (Shaxda Dheeraadka ah S3).Dhibcaha isku dhafka ah ee taxanaha tixraaca waxay u dhexeeyaan 75.5% ilaa 88.6%.Celcelis ahaan isku xidhka xogta tirinta akhrinta ee u dhaxaysa kala duwanaanshaha tijaabada ee udhaxeeya nuqullada noolaha ayaa u dhexeeyay 0.812 ilaa 0.997 (macnaha 0.959). Marka la eego 35,170 hiddo-wadaha la falanqeeyay, 2917 ma muujin wax muujin ah, iyo 4785 kale oo hidde-sidayaasha ah ayaa lagu muujiyay heer aan fiicneyn (saldhigga macnaha <5). Marka la eego 35,170 hiddo-wadaha la falanqeeyay, 2917 ma muujin wax muujin ah, iyo 4785 kale oo hidde-sidayaasha ah ayaa lagu muujiyay heer aan fiicneyn (saldhigga macnaha <5). Иz 35 170 proanalizirovannыh genov 2917 iyo kuwa kale oo badan овне (базовое среднее <5). 35,170 hiddo-wadaha la falanqeeyay, 2917 ma muujin wax hadal ah, iyo 4785 gen ee soo hadhay ayaa lagu muujiyay heer aan fiicneyn (saldhigga macnaha <5).在分析的35,170 个基因中，2917 个没有表达，其他478535,170 Иz 35 170 proanalizirovannыh genov 2917 iyo Ostalnыe 4785 е среднее значение <5). 35,170 ee hiddo-wadaha la falanqeeyay, 2917 lama sheegin halka 4785 hidde-sideyaasha soo harayna ay lahaayeen tilmaamo dayacan (saldhigga macnaha <5).Sidaa darteed, tirada hiddo-wadaha loo tixgeliyey in la muujiyey (macnaha saldhigga ≥ 5) inta lagu guda jiro tijaabada waxay ahayd 27,468 (78.1%) (Shaxda Dheeraadka ah S4).
Laga soo bilaabo barta ugu horreysa, dhammaan khadadka NLL waxay ka jawaabeen tallaalka C. lupini (cadaad Col-08) iyadoo dib loo habeeyey qoraalka qoraalka (Shaxda 1), si kastaba ha ahaatee, farqi weyn ayaa loo arkay inta u dhaxaysa xariiqyada.Sidaa darteed, xariiqda iska caabinta 83A: 476 (sida hidda-wadaha Lanr1) waxay muujisay dib-u-habayn weyn oo transcriptome ah markii ugu horreysay (6 hpi) oo leh 31-69-laab koror ah tirada hiddo-goynta-kor-iyo-hoosaadka marka la barbar dhigo dhibcaha kale ee waqtigan.Intaa waxaa dheer, kan ugu sarreeya wuxuu ahaa mid gaaban, maaddaama muujinta hiddo-wadaha dhowrka ah ay si weyn u beddeleen barta labaad (12 hpi).Waxa xiiso leh, Boregine, oo sidoo kale muujisay iska caabin heer sare ah oo ku saabsan tijaabada tallaalka, ma marin barnaamijkan qoraalka ah ee baaxadda leh inta lagu jiro tijaabada.Si kastaba ha ahaatee, tirada hiddo-sidaha si kala duwan loo muujiyay (DEG) waxay la mid ahaayeen Boregine iyo 83A: 476 ee 12 HPI.Mandelup iyo dadka 22660 labaduba waxay muujiyeen DEG ugu sarreysay markii ugu dambeysay (48 l/s), taasoo muujineysa dib u dhac yar oo ku yimid jawaabaha difaaca.
Sababtoo ah 83A: 476 waxaa lagu sameeyay dib u habeyn ballaaran oo qoraal ah iyadoo laga jawaabayo C. lupini ee 6 HPI marka la barbar dhigo dhammaan xariiqyada kale, ~ 91% DEG-yada lagu arkay waqtigan waxay ahaayeen kuwo gaar ah (Jaantus 1).Si kastaba ha ahaatee, waxaa jiray xoogaa is-dhexgal ah jawaabaha hore ee u dhexeeya khadadka daraasadda, sida 68.5%, 50.9%, iyo 52.6% DEG ee Boregine, Mandelup, iyo dadweynaha 22660, siday u kala horreeyaan, oo ay la socdaan kuwa laga helay 83A: 476 waqti cayiman.Hase yeeshee, DEG-yadani waxay ku xisaabtameen jajab yar oo keliya (0.97-1.70%) dhammaan DEG-yada hadda la ogaaday iyadoo la adeegsanayo 83A: 476.Intaa waxaa dheer, 11 DEG ee dhammaan khadadka waxay ahaayeen kuwo isku xiran waqtigan (S4-S6) oo ay ku jiraan qaybaha caadiga ah ee jawaabaha difaaca dhirta: borotiinka wareejinta dufanka (TanjilG_32225), endoglucan-1,3-β-glucoside enzyme (TanjilG_23384), laba borotiinno stress-inducible sida SAM2G_5 (TanjilG_5) borotiinka tex (TanjilG_32352), iyo laba borotiinada gidaarka qaabdhismeedka hodanka ku ah glycine (TanjilG_19701 iyo TanjilG_19702).Waxa kale oo jiray isku-xidhnaan aad u sarreeya oo jawaabaha qoraalka ah ee u dhexeeya 83A:476 iyo Boregine ee 24 HPI (wadarta 16-38% DEG) iyo inta u dhaxaysa Mandelup iyo Population 22660 ee 48 HPI (wadarta 14-20% DEG).
jaantuska Venn oo muujinaya tirada hiddo-wadaha si kala duwan loo muujiyay (DEG) ee khadadka lupine-caleemaha cidhiidhiga ah (NLL) ee lagu tallaalay Colletotrichum lupini (cadaad Col-08 laga helay beeraha lupine ee Wierzhenice, Poland, 1999).Khadadka NLL ee la falanqeeyay waxay ahaayeen: 83A: 476 (iska caabin ah, sidda Lanr1 allele), Boregine (adkeysi, asalka hidde-sidaha aan la garanayn), Mandelup (dhexdhexaad u adkaysta, sida AnMan allele) iyo dadweynaha 22660 (aad u nugul).Soo gaabinta hpi waxay taagan tahay saacado tallaalka ka dib.Qiimaha eber waa la saaray si loo fududeeyo garaafyada.
Qaybta hiddo-wadaha aadka u sarreeya ee 6 hpi ayaa lagu falanqeeyay joogitaanka xayndaabyada hidda-wadaha R ee canonical (Shaxda Dheeraadka ah S7).Daraasadani waxay daaha ka qaaday soo-dhaweynta hiddo-wadaha iska caabbinta cudurrada caadiga ah ee NBS-LRR oo keliya ee 83A: 476.Qaybtani waxay ka koobnayd hal TIR-NBS-LRR hiddo-wade (tanjilg_05042), shan CC-NBS-LRR gen (tanjilg_06165, tanjilg_06162, tanjilg_22773, tanjilg_22640, iyo tanjilg_1616-LR_4), iyo afar N1 N1 NBSLR_1 (tanjilg_16162) iyo sidoo kale Afar NBS-Lrr (tanjilg_16162) iyo Afar NBS-LRR (TANJILG_16162).Dhammaan hidde-sideyaashani waxay leeyihiin qaybo canonical ah oo loo habeeyey si isdaba joog ah.Marka laga soo tago hiddo-wadaha NBS-LRR, dhowr kinases RLL ah ayaa laga hawlgeliyay 6 hpi, kuwaas oo ah mid ku yaal Boregine (TanjilG_19877), laba ku yaal Mandelup (TanjilG_07141 iyo TanjilG_19877) iyo dadka 22660 (TanjilG_0901G) .
Hidde-yaasha leh muujinta si weyn loo beddelay oo ka jawaabaya tallaalka C. lupini ( cadaadis Col-08) ayaa lagu sammeeyay falanqaynta kobcinta Gene Ontology (GO) (Shaxda Dheeraadka ah S8). Ereyga habka bayoolojiga ee inta badan loo soo bandhigay wuxuu ahaa "GO: 0006952 jawaab celinta difaaca" kaas oo ka soo muuqday 6 ka mid ah 16 (time × line) isku darka oo leh muhiimad sare (P qiimaha <0.001) (Jaantus. 2). Ereyga habka bayoolojiga ee inta badan loo soo bandhigay wuxuu ahaa "GO: 0006952 jawaab celinta difaaca" kaas oo ka soo muuqday 6 ka mid ah 16 (time × line) isku darka oo leh muhiimad sare (P qiimaha <0.001) (Jaantus. 2). GO: 0006952 я в 6 из 16 Ereyga habka bayoolojiga ee inta badan loo soo bandhigay wuxuu ahaa 'GO: 0006952 jawaab celinta difaaca', kaas oo ka soo muuqday 6 ee 16 (time × lineage) isku darka oo leh muhiimad sare (P value <0.001) (Jaantus. 2).最常被过度代表的生物过程术语是“GO:0006952具有高显着性（P 值< 0.001）（图2）。 Ereyga habka bayoolojiga ee ugu badan ee mataleya waa "GO: 0006952 jawaabta difaaca", kaas oo ka muuqda 6 ka mid ah 16 (时间×线), oo leh muhiimad sare (P value <0.001) (图2) . "GO: 0006952 Jawaabta Difaaca" 1 комбинаций (время × линия) с высокой значимостью (значение P <0,001) (рис. 2). Ereyga habka bayoolojiga ee inta badan loo soo bandhigay wuxuu ahaa 'GO: 0006952 Jawaabta Difaaca', kaas oo ka soo muuqday 6 ka mid ah 16 isku darka (waqtiga ×) oo leh muhiimad sare (P qiimaha <0.001) (Jaantus. 2).Ereygan ayaa si xad dhaaf ah loogu soo bandhigay laba dhibcood oo 83A: 476 iyo Boregine (6 iyo 24 hpi) iyo hal mar Mandelup iyo Dadweynaha 22660 (12 iyo 6 hpi, siday u kala horreeyaan).Tani waa natiijo la filayo, oo muujinaysa jawaabta antifungal ee xadadka u adkaysta.Intaa waxaa dheer, 83A: 476 waxay ka jawaabtay C. lupini iyada oo si degdeg ah u soo jiidata hiddo-wadaha la xidhiidha dillaaca oksaydhka ee matalaya ereyga "GO: 0055114 habka redox", taas oo muujinaysa jawaab celin gaar ah oo difaac ah, halka Boregine uu muujiyay jawaabaha difaaca gaarka ah, oo la xidhiidha ereyga 'GO'.: 0006950 Ka jawaab celinta cadaadiska.Dadka 22660 waxay kiciyeen jawaab celinta iska caabbinta tooska ah ee ku lug leh dheef-shiid kiimikaad sare, taas oo muujinaysa tirada xad-dhaafka ah ee ereyada "GO: 0016104 Habka biosynthesis triterpene" iyo "GO: 0006722 Geedi socodka dheef-shiid kiimikaadka triterpene" , falcelinta hore 83A: 476 (6 hpi) iyo falcelinta daahday Mandelup iyo Population 22660 waxaa ka mid ah ereyga GO:0015979 'photosynthesis' iyo kuwa kale oo la xiriira hababka noolaha.
Erayada hiddo-wadaha bioprocess-ka ee lagu xushay sharraxaadda hiddo-wadaha si kala duwan loo muujiyey inta lagu jiro jawaabaha transcriptome ee lupine-caleemo-dhuuban (NLL) oo lagu tallaalay lupine anthrax (Col-08 oo laga helay beeraha lupine ee Wierzhenice, Poland, 1999) ayaa si weyn loo buunbuuniyay.Khadadka NLL ee la falanqeeyay waxay ahaayeen: 83A: 476 ( adkaysi u leh, siday homozygous Lanr1 allele), Boregine (u adkaysta, asalka hidde-sidaha aan la garanayn), Mandelup (dhexdhexaad u adkaysta, sidda homozygous AnMan allele) iyo dadweynaha 22660 (u nugul).
Sababtoo ah daraasaddan ayaa loogu talagalay in lagu aqoonsado hiddo-wadaha ka qaybqaata caabbinta anthracnose, hiddo-wadaha loo qoondeeyay ereyada GO "GO: 0006952 jawaabaha difaaca" iyo "GO: 0055114 hababka Redox" ayaa lagu falanqeeyay goynta tan iyo asal ahaan macnaheedu waa ≥ 30 oo leh ugu yaraan hal xariiq.× wakhtiga isku darka qiyamka tirakoobka muhiimka ah ee log2 (la laablaabashada).Tirada hidda-sidaha buuxiyey shuruudahan ayaa ahaa 65 ee GO:0006952 iyo 524 ee GO:0055114.
83A: 476 ayaa shaaca ka qaaday laba DEG ugu sarreysa oo lagu sharraxay ereyga GO: 0006952, kan ugu horreeya ee 6 hiddo-wadaha halkii inch (64 hidde-side, kor iyo hoos) iyo tan labaad oo ah 24 gen halkii inch (15 hidde, kor u kaca oo keliya).Boregine waxa kale oo ay muujisay in GO:0006952 uu ugu sarreeyey isla wakhtigaas, balse ay ka yar tahay DEG (11 iyo 8) iyo hawlgelinta door bideysa.Mandeloop wuxuu muujiyay laba heerar oo GO ah: 0006952 ee 12 iyo 48 HPI, labaduba waxay sideen 12 hiddo-wadaha (kii ugu horreeyay ee leh hiddo-wadaha firfircoon, iyo kan labaad ee leh hiddo-wadaha kaliya ee xakamaynta), halka dadka 22660 ee 6 HPI (13 gen) ay lahaayeen sare u kac weyn oo ah kororka ugu sarreeya.xeerar.Waa in la ogaadaa in 96.4% ee GO:0006952 DEG ee meelahan ugu sarreeya ay lahaayeen jawaab isku mid ah (kor ama hoos), taas oo muujinaysa isku-dhafka muhiimka ah ee jawaabaha difaaca inkasta oo ay ku kala duwan yihiin tirada hidde-sideyaasha ku lug leh.Kooxda ugu weyn ee taxanaha ah ee la xidhiidha ereyga GO: 0006952 waxay qeexaysaa Farriinta Cadaadiska Gaajada-Associated Message Protein 22 (SAM22-like), kaas oo iska leh fasalka 10 ee borotiinka-pathogenesis-ku xiran borotiinka (PR-10) borotiinka clade iyo borotiinka asaasiga ah.borotiin la mid ah (MLP-sida) borotiin (Jaantus 3).Labada kooxood waxay ku kala duwanaayeen dabeecadda hadalka iyo jihada jawaabta.Hidde-sidayaasha ku-meel-gaadhka ah ee SAM22-sida borotiinada waxay muujiyeen soo-galin joogto ah oo muhiim ah waqtiyadii hore (6 ama 12 hpi) waxayna ahaayeen kuwo aan ka jawaabin dhammaadka tijaabada (48 hpi), halka borotiinnada MLP-u eg ay muujiyeen isku-duwidda 6 hpi.hpi.83A: 476 iyo Mandelup ee 48 hp/in, ku dhawaad ​​dhammaan dhibcooyinka kale ee xogtu ma jawaabin.Intaa waxaa dheer, kala duwanaanshaha astaamaha muujinta ee hiddo-wadaha borotiinka ee SAM22 ayaa raacay kala duwanaanshiyaha iska caabbinta anthracnose, maadaama khadadka u adkaysiga badan ay heleen dhibco waqti badan oo si weyn u soo jiidaya hiddo-wadahaas marka loo eego gen-yo nugul.Hidde-sidayaasha kale ee LlR18A/B-u eg PR-10 ayaa muujiyay hannaanka muujinta la midka ah ee hiddasidaha borotiinka ee SAM22.
Qaybaha ugu muhiimsan ee habka bayoolojiga ereyga "GO: 0006952 Jawaabta Difaaca" iyo qaababka muujinta hiddo-wadaha musharaxiinta ee Lanr1 iyo AnMan alleles ayaa la aqoonsaday.Miisaanka Log2 wuxuu u taagan yahay qiyamka log2 (isbeddel laablaab) inta u dhaxaysa la tallaalay (Colletotrichum lupini, cadaadis Col-08, oo laga helay beeraha lupine, Wizhenica, Poland, 1999) iyo kontoroolka (sham-ku-tallaalidda) dhirta isla mar ahaantaana.Khadadka lupine-ka cidhiidhiga ah ee soo socda ayaa la falanqeeyay: 83A: 476 ( adkaysi u leh, sidda homozygous Lanr1 allele ), Boregine ( adkaysi u leh, asalka hidde-sidaha aan la garanayn), Mandelup (dhexdhexaad u adkaysta, sidda homozygous AnMan allele), iyo Dadweynaha 22660 (u nugul).
Intaa waxaa dheer, muuqaalada muujinta RNA-seq genes musharaxa Lanr1 (TanjilG_05042) iyo AnMan (TanjilG_12861) ayaa la qiimeeyay (Jaantus. 3).Hidde-sidaha TanjilG_05042 wuxuu muujiyay jawaab celin muhiim ah (dhaqdhaqaaq) 83A: 476 kaliya marka ugu horeysa (6 hpi), halka TanjilG_12861 uu muhiim u ahaa Mandeloop kaliya laba dhibcood: 6 hpi (sharciga hoose) iyo 24 hpi (6 hpi).leh.).la hagaajin karo)).
Hidde-sidayaasha aadka u xad-dhaafka ah ee ereyga GO: 0055114 "habraaca dib-u-celinta" waxay ahaayeen hidde-socodyada cytochrome P450 borotiinada iyo peroxidase (Jaantus. 4).Shaybaarada laga soocay 83A: 476 at 6 HPI, ugu badnaan ama ugu yaraan log2 (laab beddelka isbeddelka) qiyamka (86.6% ee hiddo-wadaha) ayaa guud ahaan lagu arkay inta u dhaxaysa dhirta la tallaalay iyo kuwa la kantaroolo, taas oo muujinaysa jawaabta sare ee genotype this ee tallaalka galmada.83A: 476 waxay muujisay GO-ga ugu muhiimsan: 0055114 DEG at 6 hpi (503 genes), halka inta soo hartay ee 48 hpi (Boregine, 31 genes; Mandelup, 85 genes; iyo Population 22660, 78 genes)).Inta badan hiddo-wadaha GO: 0055114 qoyska, laba nooc oo jawaab celin ah oo tallaalka ah (firfircooni iyo xannibaad) ayaa lagu arkay.Waxa xiiso leh, ilaa 97.6% DEG-yada loo aqoonsaday ereyga GO: 0055114 ee Mandelupe ee 48 hp Aragtiyadani waxay soo jeedinayaan in, inkastoo miisaan aad u yar (ie, tirada hiddo-wadaha redox-ka ee la beddelay, 85 oo ka soo horjeeda 503), qaabka dib u dhigista jawaabaha transcriptome ee manracnose4 ayaa la mid ah jawaab-celinta 6.Boregine iyo Dadweynaha 22660, isku-darkani wuu hooseeyaa 51.6% iyo 75.6%, siday u kala horreeyaan.
Nidaamyada muujinta qaybaha ugu muhiimsan ee ereyga geeddi-socodka bayoolojiga "GO: 0055114 habka dib u habeynta" ayaa la muujiyay.Miisaanka Log2 wuxuu u taagan yahay qiyamka log2 (isbeddel laablaab) inta u dhaxaysa la tallaalay (Colletotrichum lupini, cadaadis Col-08, oo laga helay beeraha lupine, Wizhenica, Poland, 1999) iyo kontoroolka (sham-ku-tallaalidda) dhirta isla mar ahaantaana.Khadadka lupine-ka cidhiidhiga ah ee soo socda ayaa la falanqeeyay: 83A: 476 ( adkaysi u leh, sidda homozygous Lanr1 allele ), Boregine ( adkaysi u leh, asalka hidde-sidaha aan la garanayn), Mandelup (dhexdhexaad u adkaysta, sidda homozygous AnMan allele), iyo Dadweynaha 22660 (u nugul).
83A: 476 Jawaabaha transcriptomic ee tallaalka C. lupini (cadaad Col-08) sidoo kale waxaa ku jiray aamusnaanta isku-duwidda ee hiddo-wadaha loo aaneeyay ereyga GO: 0015979 "photosynthesis" iyo hababka kale ee bayoolojiga la xiriira (FIG. 5).Qaybtan GO:0015979 DEG waxay ka koobnayd 105 hidde-side oo si wayn loo cadaadiyay 6 hpi at 83A:476.Qaybtan hoose, 37 hiddo-wadaha ayaa sidoo kale hoos loo dhigay Mandelup ee 48 HPI iyo 35 isla mar ahaantaana dadka 22660, oo ay ku jiraan 19 DEGs oo caadi u ah labada genotypes.Ma jiro DEGs oo la xidhiidha ereyga GO: 0015979 si aad ah ayaa loo hawlgeliyay marka la isku daro (wakhtiga x).
Qaababka muujinta qaybaha ugu muhiimsan ee ereyga geeddi-socodka noolaha "GO: 0015979 Photosynthesis" ayaa la muujiyay.Miisaanka Log2 wuxuu u taagan yahay qiyamka log2 (isbeddel laablaab) inta u dhaxaysa la tallaalay (Colletotrichum lupini, cadaadis Col-08, oo laga helay beeraha lupine, Wizhenica, Poland, 1999) iyo kontoroolka (sham-ku-tallaalidda) dhirta isla mar ahaantaana.Khadadka lupine-ka cidhiidhiga ah ee soo socda ayaa la falanqeeyay: 83A: 476 ( adkaysi u leh, sidda homozygous Lanr1 allele ), Boregine ( adkaysi u leh, asalka hidde-sidaha aan la garanayn), Mandelup (dhexdhexaad u adkaysta, sidda homozygous AnMan allele), iyo Dadweynaha 22660 (u nugul).
Iyada oo ku saleysan natiijooyinka falanqaynta muujinta kala duwanaanta iyo sida loo maleynayo in ay ku lug leedahay jawaabaha difaaca ee fangaska pathogenic, shaxdan toddobo hiddo-wade ayaa loo xushay qiyaasidda astaamaha muujinta PCR-da dhabta ah (Shaxda Dheeraadka ah S9).
Hidde-sidaha borotiinka la saaray ee TanjilG_10657 ayaa si weyn loogu soo jiiday dhammaan khadadka la darsay iyo dhibcaha waqtiga marka la barbar dhigo kontoroolka (mimic) dhirta (Shaxda Dheeraadka ah S10, S11).Intaa waxaa dheer, astaanta muujinta ee TanjilG_10657 waxay muujisay isbeddel sii kordhaya muddada tijaabada ee dhammaan khadadka.Dadka 22660 waxay muujiyeen dareenka ugu sarreeya ee TanjilG_10657 si loogu tallaalo firfircoonida 114-laab iyo heerka ugu sarreeya ee muujinta qaraabada (4.4 ± 0.4) ee 24 HPI (Jaantus. 6a).Hidde-sidaha borotiinka PR10 LlR18A TanjilG_27015 waxa kale oo uu muujiyey firfircoonida dhammaan xariiqyada iyo dhibcaha wakhtiga, oo leh muhiimad tira koob oo ah dhibcaha xogta (Jaantus. 6b).Si la mid ah TanjilG_10657, heerka ugu sarreeya qaraabada ee TanjilG_27015 ayaa lagu arkay 22660 dadka la tallaalay ee 24 HPI (19.5 ± 2.4).Aashitada endochitinase hidda-wadaha TanjilG_04706 ayaa si weyn kor loogu qaaday dhammaan saadka iyo waqti kasta marka laga reebo Boregine 6 hpi (Jaantus. 6c).Waxaa si xooggan loo kiciyay barta ugu horreysa (6 HPI) ee 83A: 476 (10.5 jeer) oo si dhexdhexaad ah ayaa loo kordhiyay khadadka kale (6.6-7.5 jeer).Inta lagu jiro tijaabada, muujinta TanjilG_04706 ayaa ku hadhay heerar la mid ah 83A: 476 iyo Boregine, halka Mandelup iyo Population 22660 ay si weyn u korodhay, gaarey qiyamka sare (5.9 ± 1.5 iyo 6.2 ± 1.5, siday u kala horreeyaan).endoglucan-1,3-β-glucosidase-sida hidda-wadaha TanjilG_23384 ayaa muujiyay firfircooni sare ee labada dhibcood ee ugu horreeya (6 iyo 12 hpi) dhammaan khadadka marka laga reebo dadweynaha 22660 (Jaantus. 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).Markay tahay 24 HPI, TanjilG_23384 muujinta waxay ahayd mid aad u hooseeya dhammaan khadadka la bartay (laga bilaabo 0.04 ± 0.009 ilaa 0.44 ± 0.12).
Tilmaamaha muujinta hiddo-wadaha la xushay (ag) oo ay daaha ka qaaday tirada PCR.Lambarada 6, 12 iyo 24 waxay matalaan saacado ka dib tallaalka.Hidde-sideyaasha LanDExH7 iyo LanTUB6 ayaa loo isticmaalay si caadi ah iyo LanTUB6 waxa loo isticmaalay isku-habaynta taxanaha.Baararka khaladku waxa ay u taagan yihiin weecista caadiga ah ee ku salaysan saddex nuqul oo bayooloji ah, kuwaas oo mid walba uu yahay celceliska saddex nuqul oo farsamo. Muhiimadda tirakoobka ee kala duwanaanshaha heerarka tibaaxaha ee u dhexeeya la tallaalay (Colletotrichum lupini, cadaadis Col-08, laga helay 1999 laga soo bilaabo beerta lupine ee Wierzenica, Poland) iyo kantaroolka (majaajilada-macnaha) dhirta ayaa lagu calaamadeeyay dhibcaha xogta (* Qiimaha P <0.05, *** P qiimaha ≤ 0.01). Muhiimadda tirakoobka ee kala duwanaanshaha heerarka tibaaxaha ee u dhexeeya la tallaalay (Colletotrichum lupini, cadaadis Col-08, laga helay 1999 laga soo bilaabo beerta lupine ee Wierzenica, Poland) iyo kantaroolka (majaajilada-macnaha) dhirta ayaa lagu calaamadeeyay dhibcaha xogta (* Qiimaha P <0.05, *** P qiimaha ≤ 0.01). Статическая значимость различий уровнях эkspressы mezhdu iyokulyirovannыmy люпина в Верженице, Польша) iyo контрольными P ≤ 0,01, ***значение P ≤ 0,001). Tirakoob ahaan farqiga u dhexeeya heerarka muujinta ee u dhexeeya la tallaalay (Colletotrichum lupini, cadaadis Col-08, laga helay 1999 laga soo bilaabo beerta lupine ee Wierzhenice, Poland) iyo xakamaynta (sham-inocated) dhirta ayaa lagu xusay dhibcaha xogta (* Qiimaha <0.05, ** P-qiimaha ≤ 0.01, ***-0).接种（Colletotrichum lupini，Col-08株，1999年从波兰Wierzenica的羽扇豆田获得）和对照（模拟接科（模拟接科剩模拟接科前模拟接科剤來來來）差异的统计学显着性标记在数据点上方（*P值< 0.05, **P 值≤ 0.01, ***P 值≤ 0.001).接种 （colletotrichum lupini, color-08 株 ， 1999 年 波兰 波兰 wierzenica差异 的 统计学 显着性 标记 数据点 上方*p 值 <0.05, **P ≤ 0.01, ***P ≤ 0.001). STATICYCHESKI ZNACHIMY RAZILICIA V UREVNYAH эkspressi mezhdu ynokulirovannymi Верженице, Польша, в 1999 г.) iyo контрольными. -значение ≤ 0,01, ***P-значение ≤ 0,001). Tirakoob ahaan farqiga u dhexeeya heerarka muujinta ee u dhexeeya la tallaalay (Colletotrichum lupini, cadaadis Col-08, laga helay beeraha lupine ee Verzhenice, Poland, 1999) iyo xakamaynta (sham-inocated) dhirta ayaa lagu xusay kor ku xusan dhibcaha xogta (* Qiimaha P <0.05, ** P-qiimaha ≤ 0.01, *** P-01).Khadadka NLL ee la falanqeeyay waxay ahaayeen: 83A: 476 ( adkaysi u leh, oo sita homozygous Lanr1 allele), Mandelup (dhexdhexaad u adkaysta, oo sita homozygous AnMan allele), Boregine (u adkaysta, asalka hidde-sidaha aan la garanayn) iyo dadweynaha 22660 (u nugul).
Hinsidaha musharaxa TanjilG_05042 ee goobta Lanr1 wuxuu muujiyay qaab muujineed si muuqata uga duwan muuqaalada laga helay daraasadaha RNA-seq (Jaantus. 6e).Dhaqdhaqaaqa muhiimka ah ee hidde-sidahan ayaa lagu arkay Mandelup iyo dadka 22660 (illaa 39.7 iyo 11.7, siday u kala horreeyaan), taasoo keentay heerar muujin oo sarreeya (ilaa 1.4 ± 0.14 iyo 7.2 ± 1.3, siday u kala horreeyaan).83A: 476 ayaa sidoo kale shaaca ka qaaday qaar ka mid ah kor u qaadista hidda-wadaha TanjilG_05042 (ilaa 3.8-laab), si kastaba ha ahaatee, heerarka muujinta qaraabada ah ee la gaarey (0.044 ± 0.002) waxay ka badan yihiin 30-laab oo ka hooseeya kuwa lagu arkay Mandelup iyo 22660 dadweynaha.Falanqaynta qPCR waxay muujisay farqi weyn oo u dhexeeya heerarka odhaahda genotypes ee noocyada la tallaalay (xakamaynta), taasoo gaadhay farqi 58 laab ah oo u dhexeeya dadka 22660 iyo 83A: 476, iyo sidoo kale inta u dhaxaysa dadka 22660 iyo 22660. Farqi laba laab ah ayaa la gaadhay inta u dhaxaysa Boregine iyo Mandalup.
Hidde-sidaha musharaxa ee goobta AnMan, TanjilG_12861, ayaa la hawlgeliyay iyada oo laga jawaabayo tallaalka 83A: 476 iyo Mandelup, wuxuu ahaa dhexdhexaad dadka 22660, waxaana hoos loo dhigay Boregine (Jaantus. 6f).Muujinta qaraabada ah ee hiddo-wadaha TanjilG_12861 ayaa ahaa kan ugu sarreeya ee la tallaalay 83A: 476 (0.14± 0.01).Heerka 17.4 kDa I hiddo-wadaha borotiinka naxdinta leh ee TanjilG_05080 HSP17.4 ayaa muujiyay heerarka muujinta qaraabada hoose ee dhammaan noocyada iyo dhibcaha waqtiga ee la bartay (Jaantus. 6g).Qiimaha ugu sarreeya ayaa lagu arkay 24 HPI ee dadweynaha 22660 (0.14 ± 0.02, koror siddeed laab ah oo ka jawaabaya tallaalka).
Isbarbardhigga astaamaha muujinta hidde-sidaha (Jaantus. 7) ayaa muujiyay xidhiidh sare oo u dhexeeya TanjilG_10657 iyo afar hidde-sideyaal kale: TanjilG_27015 (r = 0.89), TanjilG_05080 (r = 0.85), TanjilG_05042 (r = 0.80) (r = 0.80), iyo Tanjil.Natiijooyinka noocan oo kale ah ayaa laga yaabaa inay muujiyaan isku-darka gen ee kuwan inta lagu jiro jawaabaha difaaca.Hidde-sidayaasha TanjilG_12861 iyo TanjilG_23384 waxay muujiyeen muuqaalo muujineed kala duwan oo leh qiimayaal isku xidhid hoose oo Pearson ah (laga bilaabo 0.08 ilaa 0.43 iyo -0.19 ilaa 0.28, siday u kala horreeyaan) marka la barbar dhigo hiddo-wadaha kale.
Xidhiidhka u dhexeeya astaamaha muujinta hidda-wadaha ayaa lagu ogaaday iyadoo la isticmaalayo PCR-ga tirada leh.Khadadka lupine-ka cidhiidhiga ah ee soo socda ayaa la falanqeeyay: 83A: 476 ( adkaysi u leh, oo sita homozygous Lanr1 allele), Mandelup (dhexdhexaad u adkaysta, sidda homozygous AnMan allele), Boregine ( adkaysi, asalka hidde aan la garanayn), iyo Dadweynaha 22660 (u nugul).Saddexda dhibcood ee wakhtiga ayaa la xisaabiyay (6, 12 iyo 24 saacadood ka dib tallaalka), oo ay ku jiraan la tallaalay (Colletotrichum lupini, strain Col-08, oo laga helay beeraha lupine ee Wierzhenice, Poland, 1999) iyo xakamaynta (sham-inocated) dhirta.Miisaanka ayaa muujinaya qiimaha isku xidhka Pearson.
Iyada oo ku saleysan xogta laga helay 6 hooras halkii inch, WGCNA waxaa lagu sameeyay 9981 DEG oo lagu aqoonsaday iyadoo la is barbar dhigayo dhirta la tallaalay iyo tan la xakameeyey si diiradda loo saaro jawaabaha difaaca hore (Sanduuqa Dheeraadka ah S12).Laba iyo labaatan nooc oo hiddo-wadaha ah (kutlada) ayaa lagu helay muuqaallo isku xidhan (togan ama taban) oo u dhexeeya noocyada genotypes iyo kala duwanaanshaha tijaabada ah. Celcelis ahaan, heerarka muujinta hidde-sidaha ayaa hoos u dhacay si ay u kala horreeyaan 83A: 476> Mandelup> Boregine> Dadka 22660 (labada nooc, si kastaba ha ahaatee, isbeddelkani wuxuu ahaa mid ka xoog badan dhirta xakamaynta). Celcelis ahaan, heerarka muujinta hidde-sidaha ayaa hoos u dhacay si ay u kala horreeyaan 83A: 476> Mandelup> Boregine> Dadka 22660 (labada nooc, si kastaba ha ahaatee, isbeddelkani wuxuu ahaa mid ka xoog badan dhirta xakamaynta). 83A: 476 > Mandelup > Boregine > Dadka 22660 ильнее у контрольных растений). Celcelis ahaan, heerarka muujinta hidde-sidaha ayaa hoos u dhacay sida ay u kala horreeyaan 83A: 476> Mandelup> Boregine> Dadka 22660 (labada nooc, si kastaba ha ahaatee, isbeddelkani wuxuu ahaa mid ka xoog badan dhirta xakamaynta).平均而言，基因表达水平按83A:476 > Mandelup > Boregine > Population 22660物中更强）平均 而 言 ， 基因 水平 按 按 83a: 476> mandelup> boregine> dadka 22660 . .............. 83A: 476 > Mandelup > Boregine > Dadka 22660 е у контрольных растений). Celcelis ahaan, heerarka muujinta hidde-sidaha ayaa hoos u dhacay taxanaha 83A: 476> Mandelup> Boregine> Dadka 22660 (si kastaba ha ahaatee, labada noocba, isbeddelkani wuxuu ahaa mid ka xoog badan dhirta xakamaynta).Tallaalku waxa uu keenay kor u qaadista muujinta hidde-sidaha, gaar ahaan cutubyada 18, 19, 14, 6 iyo 1 (oo ah siday u kala horreeyaan), nidaam taban (tusaale cutubyada 9 iyo 20) ama saamayn dhexdhexaad ah (tusaale cutubyada 11, 22, 8 iyo 13).Falanqaynta kobcinta ereyga GO (Shaxda Dheeraadka ah S13) ayaa shaaca ka qaaday "GO: 0006952 jawaabaha ilaalinta" ee moduleka la tallaalay (18) oo leh firfircooni ugu badan, oo ay ku jiraan hiddo-wadaha lagu falanqeeyay qPCR modules (9).Module 18 concentrator (Jaantus. 8) ayaa loo aqoonsaday inuu yahay hidda-wadaha TanjilG_26536 oo codaynaya borotiinka PR-10-sida LlR18B, iyo module 9 concentrator waxaa loo aqoonsaday inuu yahay TanjilG_28955 hiddasidaha codaynta nidaamka sawirka II PsbQ borotiinka.Musharaxa caabbinta anthracnose hidda-wadaha Lanr1, TanjilG_05042, ayaa laga helay moduleka 22 (Jaantus. 9) wuxuuna la xidhiidhaa ereyada "GO: 0044260 hababka dheef-shiid kiimikaadka unugyada unugyada" iyo "GO: 0006355 Xeerka qoraalka, DNA templating" oo sita Hubka TanjilG_0121Hidde-siduhu waxa uu dhigayaa qodobka qoraalka cadaadiska kulaylka A-4a (HSFA4a).
Falanqaynta shabakada miisaanka leh ee wada-shaqeynta hiddo-wadaha cutubyada leh ereyada habka bayoolojiga ee aad loo soo bandhigay "GO: 0006952 jawaabaha difaaca".Ligation ayaa la fududeeyay si loo muujiyo afarta hidde ee uu falanqeeyay qPCR (TanjilG_04706, TanjilG_23384, TanjilG_10657 iyo TanjilG_27015).
Falanqaynta shabakada miisaanka leh ee wada-shaqeynta hidda-wadaha moduleka leh habka bayooloji ee si xad dhaaf ah loo soo bandhigay "GO: 0006355: Xeerarka qoraalka, qaabaynta DNA" iyo sidda musharaxa hidda-socodka iska caabinta anthracnose Lanr1 TanjilG_05042.Liistada ayaa la fududeeyay si loo karantiilo hiddo-wadaha TanjilG_05042 iyo hiddo-wadaha TanjilG_01212.
Baaritaanka iska caabbinta Anthracnose ee laga soo aruuriyay Australia ayaa muujisay in badi beerihii hore ee la sii daayay ay u nugul yihiin;Kalya, Coromup iyo Mandelup ayaa lagu tilmaamay kuwo dhexdhexaad ah u adkaysta, halka Wonga, Tanjil iyo 83A:476 lagu tilmaamay inay yihiin kuwo aad u adkaysi badan26,27,31.waxay lahaayeen allele isku mid ah oo iska caabin ah, oo loo qoondeeyay Lanr1, iyo Coromup iyo Mandelup waxay lahaayeen allele ka duwan, oo loo qoondeeyay AnMan10, 26, 39, halka Kalya ay dhaaftay allele kale., Lanr2.Baaritaanka caabbinta anthracnose ee Jarmalka waxay keentay aqoonsiga xariiqda iska caabinta Bo7212 ee leh musharax allele aan ahayn Lanr1, oo loo qoondeeyay LanrBo36.
Daraasaddeennu waxay daaha ka qaaday soo noqnoqoshada aad u hooseeya (qiyaastii 6%) ee Lanr1 allele ee jeermiska la tijaabiyay.U fiirsashadani waxay la socotaa natiijooyinka baaritaanka jeermiska Bariga Yurub iyadoo la adeegsanayo calaamadaha Anseq3 iyo Anseq4, taas oo muujisay in Lanr1 allele uu ku jiro laba xariiq oo Belarusiyaan ah.Tani waxay soo jeedinaysaa in Lanr1 allele aan weli si weyn loo isticmaalin barnaamijyada taranka maxalliga ah, si ka duwan Australia, halkaas oo ay tahay mid ka mid ah alleles muhiimka ah ee taranta-caawinaysa taranta.Tan waxa laga yaabaa inay sabab u tahay heerka hoose ee iska caabinta ay bixiso Lanr1 allele ee xaaladaha gudaha Yurub marka la barbar dhigo warbixinta Australia.Intaa waxaa dheer, daraasaadka kudka anthracnose ee meelaha roobabka badan ee Australia waxay muujiyeen in jawaabaha iska caabinta ee ay dhexdhexaadinayaan Lanr1 allele laga yaabo inaysan waxtar u lahayn xaaladaha cimilada ee door bida korriinka iyo horumarka degdega ah ee pathogen19,42.Dhab ahaantii, daraasaddan hadda jirta, calaamadaha qaar ee anthracnose ayaa sidoo kale lagu arkay genotypes oo sita Lanr1 allele, taas oo soo jeedinaysa in iska caabintu ay meesha ka baxdo xaaladaha ugu wanaagsan ee horumarinta C. lupini.Intaa waxaa dheer, fasiraadaha been abuurka ah ee joogitaanka calaamadaha Anseq3 iyo Anseq4, kuwaas oo qiyaastii 1 cm ka yimid goobta Lanr1, ayaa suurtagal ah 28,30,43.
Daraasaddeenu waxay muujisay in 83A: 476, oo qaadaya Lanr1 allele, waxay ka jawaabtay tallaalka C. lupini oo leh dib-u-habeeyn ballaaran oo transcriptome ah oo ku saabsan wakhtiga ugu horreeya ee la falanqeeyay (6 hpi), halka Mandelup, oo xambaarsan AnMan allele, jawaabaha transcriptomic ayaa la arkay wax badan ka dib.(laga bilaabo 24 ilaa 48 hp).Kala duwanaanshahan ku-meel-gaadhka ah ee jawaabaha difaaca ayaa la xidhiidha kala duwanaanshaha calaamadaha cudurka, taas oo muujinaysa muhiimada hore ee aqoonsiga pathogene ee jawaab-celinta guusha leh ee caabbinta.Si ay u faafiyaan nudaha geedka, kudka kudka waa in ay maraan dhowr marxaladood oo korriin ah oo ku yaala dulka martida loo yahay, oo ay ku jiraan biqilka, kala qaybinta unugyada, iyo samaynta appressorium.Lifaaqa waa qaab-dhismeed aan fiicneyn oo ku dheggan dusha sare ee martida loo yahay oo sahlaysa dhexgalka unugyada martida loo yahay.Sidaa daraadeed, xadhkaha C. gloeosporioides ee soosaaray digirta ayaa muujiyay qaybta koowaad ee nukleus ka dib 75-90 daqiiqo ka dib, samaynta tuubada jeermiska ka dib 90-120 daqiiqo, iyo xakamaynta ka dib saacadaha 4 45.Mango C. gloeosporioides waxay muujisay in ka badan 40% biqilka conidial 3 saacadood ka dib iyo qiyaastii 20% samaynta cadaadiska 4 saacadood ka dib.CAP20 hiddo-wadaha fayraska-ku-xiran ee C. gloeosporioides ayaa muujiyay waxqabadyo qoraal ah oo ku saabsan epiphyte-forming conidia ka dib markii 3.5 h ku-dhufashada ee dusha sare ee avocado oo leh qadar sare oo ah borotiinka CAP20 ka dib 4 h 46 min.Sidoo kale, waxqabadka melanin biosynthesis gen ee C. trifolii ayaa la kiciyay inta lagu guda jiro 2-saac oo ku-meel-gaar ah oo ay ku xigto samaynta appressorium ka dib 1 saac.Daraasado lagu sameeyay unugyada caleenta ayaa muujiyay in strawberries lagu tallaalay C. acutatum ay marka hore xakamaynayeen 8 hpi, halka tamaandhada lagu tallaalay C. coccodes ay markii ugu horeysay xakamaynayeen 4 hpi48,49.si weyn u waafaqsan qiyaasta wakhtiga ee Colletotrichum spp.habka faafa.Jawaabaha difaaca degdega ah ee 83A: 476 waxay soo jeedinayaan ku lug lahaanshaha iska caabinta dhirta iyo hiddo-wadeyaasha difaac-kicinta (ETI) ee xariiqan, halka jawaabaha daahitaanka Mandelup ay taageerayaan xasaanadda qaab-dhismeedka molecular-ku-xiran-yar ee-kicinta (MTI) hypothesis 50. Jawaabaha hore ee 83A: 476 iyo MandelIsku-dhafka qaybeed ee u dhexeeya hiddo-wadaha kor u kaca-ama hoos-u-dejinta ee jawaab-celinta dib-u-dhacku waxay sidoo kale taageertaa fikraddan, maaddaama ETI inta badan loo tixgeliyo inay tahay jawaab-celin degdeg ah oo la xoojiyey oo MTI ah oo ku dhammaata dhimashada unugyada barnaamijka ee goobta caabuqa, oo loo yaqaan shoogga anaphylactic 51,52.
Inta badan hiddo-wadaha loo nisbeeyo ereyga xad-dhaafka ah ee Gene Ontology GO: 0006952 "Jawaabta Difaaca" waa 11 homologues ee farriinta soonka ee walaaca ay keentay 22 borotiin (oo la mid ah SAM22) iyo toddobada borotiinka-jecel ee ugu weyn (MLPs).-sida borotiinada 31, 34, 43 iyo 423 waxay muujiyeen isku xigxiga.Hidde-sidayaasha-sida SAM22 waxay muujiyeen firfircooni muhiim ah oo socday muddo dheer, taas oo muujinaysa heerarka korodhka caabbinta anthracnose (83A: 476 iyo Boregine).Si kastaba ha ahaatee, hiddo-wadaha MLP-u-eg ayaa hoos loo dhigay oo keliya xariiqyada sidda musharaxa iska caabinta allele (83A:476/Lanr1 at 6 hpi iyo Mandelup/AnMan at 24 hpi).Waa in la ogaadaa in dhammaan noocyada loo yaqaan 'SAM22-like homologues' ay asal ahaan ka soo jeedaan kooxda hidde-sideyaasha oo dhan qiyaastii 105 kb, halka hidde-sideyaasha MLP-ga ay ka soo jeedaan gobollo kala duwan oo genome ah.Dhaqdhaqaaqa isku-dhafka ah ee SAM22-sida hiddo-wadaha ayaa sidoo kale laga helay daraasaddeennii hore ee NLL caabbinta tallaalka Diaporthetoxica, oo soo jeedinaysa inay ku lug leeyihiin qaybaha jiifka ee jawaabta difaaca.Gabagabadaan waxaa sidoo kale lagu taageeray warbixinada jawaabta togan ee hiddo-wadaha SAM22-sida dhaawaca ama daaweynta salicylic acid, fangaska inducers, ama hydrogen peroxide.
Hidde-sideyaasha MLP-sida ayaa la muujiyay inay ka jawaabaan walaacyada abiotic iyo biotic kala duwan, oo ay ku jiraan bakteeriyada, fayrasyada iyo caabuqyada fangas ee pathogenic ee noocyo badan oo dhirta55.Tilmaamaha ka jawaabista isdhexgalka qaarkood ee ka dhexeeya dhirta iyo cudur-sidaha waxay u dhexeeyaan si xoog leh u kordheysa (ie, inta lagu jiro suufka Verticillium dahliae) oo si weyn hoos ugu dhacay (tusaale, ka dib caabuqa geedka tufaaxa ee Alternaria spp.)56,57.Hoos-u-dhac weyn oo MLP-sida 423 hiddo-wadaha ayaa la arkay intii lagu jiray difaaca avocado ee F. niger iyo inta lagu jiro caabuqa geedka tufaaxa Botryosphaeria berengeriana f.cn.piricola iyo Alternaria alternata waa tufaax pathotypes58,59.Intaa waxaa dheer, tufaax calli oo si xad dhaaf ah u muujinaya hidda-wadaha MLP-sida 423 ayaa lahaa muujin hoose oo ah hiddo-wadaha la-xidhiidha iska caabinta waxayna aad ugu nugul yihiin caabuqa fangas59.Ka dib Fusarium oxysporum f, hiddo-wadaha MLP-sida 423 ayaa sidoo kale lagu xakameeyay jeermiska bean caadiga ah ee adkaysi u leh.cn.Caabuqa bean 60.
Xubnaha kale ee qoyska PR-10 ee lagu aqoonsaday daraasaddeena RNA-seq waxay ahaayeen hiddo-wadaha LlR18A iyo LlR18B iyagoo ka jawaabaya kor u kaca, iyo sidoo kale hidda-wadaha la hagaajiyay (1 hiddo-wadaha).Intaa waxaa dheer, WGCNA waxay iftiiminaysaa hidda-wadaha LlR18B inay xuddun u tahay cutubkan, kaasoo aad ugu nugul tallaalka oo sidda dhowr hidde-side jawaab celin ah.LlR18A iyo LlR18B hiddo-wadaha ayaa lagu soo rogay caleemaha lupine ee jaalaha ah si ay uga jawaabaan bakteeriyada pathogenic, iyo sidoo kale NLL afkoda ka dib D. toxica inoculation, halka homologue bariiska ee hiddo-wadayaasha, RSOsPR10, ayaa si degdeg ah u keentay infekshanka fangas loo maleynayo in ay ku lug leedahay jasmonic acid signaling pathway IR16,62. s kuwaas oo looga baahan yahay bilawga caabbinta habaysan ee la helay (SAR).Iyadoo horumarinta falcelinta difaaca, borotiinka DIR1 waxaa laga soo qaadaa diiradda caabuqa iyada oo loo marayo phloem si loo kiciyo SAR xubnaha fog.Waxa xiiso leh, hiddo-wadaha TanjilG_02313 DIR1 ayaa markii ugu horreysay si weyn loogu soo jiiday sadarrada 84A: 476 iyo Dadweynaha 22660, laakiin iska caabbinta anthracnose ayaa si guul leh u soo baxday oo keliya sadarka 84A: 476.Tani waxay muujin kartaa qayb-hoosaadyo-hoosaadyada hiddo-wadaha DIR1 ee NLL, maadaama saddexda homologues ee soo hadhay ay ka jawaabeen tallaalka oo keliya 83A: 476 xariiqda 6 hpi, jawaabtana waxaa loo hagayay hoos.
Daraasaddeena, qaybaha ugu caansan 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-yagu waxay qeexaysaa HSFA4a homologue inay tahay xudun qaadata cutubyo sida musharaxa hidde-sidaha Lanr1 TanjilG_05042.HSFA4a waa qayb ka mid ah xeerka ku-tiirsanaanta redox ee qorista nukliyeerka ee dhirta.
Cytochrome P450 borotiinada waa oxidoreductases kuwaas oo kicinaya NADPH iyo / ama O2-ku-tiirsanaanta hydroxylation ee dheef-shiid kiimikaad hoose iyo sare, oo ay ku jiraan dheef-shiid kiimikaadka xenobiotics, iyo sidoo kale hormoonnada, acids fatty acids, sterols, qaybaha gidaarka unugyada, bioopolymers, iyo biosynthesis ee xeryahooda ilaalinta 69. 6 log2 (isbeddel laablaab) ilaa 5.7 sababtoo ah tiro badan oo ah homologues la beddelay (37) iyo kala duwanaanshiyaha qaababka jawaabta ee u dhexeeya hiddo-wadaha gaarka ah, taasoo ka tarjumaysa dib u eegis sare..Isticmaalka xogta RNA-seq kaliya si loo caddeeyo shaqada bayoolojiga ee gen ee NLL ee borotiinka weyn ee qoyska ayaa noqon doona mid mala-awaal ah.Si kastaba ha ahaatee, waxaa xusid mudan in qaar ka mid ah hiddo-wadaha cytochrome P450 ay la xiriiraan kororka caabbinta fangaska pathogenic ama bakteeriyada, oo ay ku jiraan wax ku biirinta falcelinta xasaasiyadda69,70,71.
Heerka III peroxidases waa enzymes dhirta kala duwan oo ku lug leh noocyo kala duwan oo hababka dheef-shiid kiimikaadka inta lagu jiro koritaanka iyo horumarinta dhirta, iyo sidoo kale ka jawaab celinta cadaadiska deegaanka sida salinity, abaarta, iftiinka iftiinka sare, iyo weerarka pathogen72.Peroxidases waxay ku lug leeyihiin isdhexgalka dhowr nooc oo dhir ah oo 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. lagenarium73,764.75Jawaabtu waa mid aad u degdeg badan, mararka qaarkood xitaa 4 HPI, ka hor inta aan fungusku soo gelin unugyada dhirta73.Hidde-sidaha peroxidase ayaa sidoo kale ka jawaabay tallaalka D. toxica NLL.Marka lagu daro shaqooyinkooda caadiga ah si ay u xakameeyaan dilaaca oksaydhka ama baabi'inta cadaadiska oksaydhka, peroxidases waxay faragelin karaan koritaanka pathogene iyagoo abuuraya caqabado jireed oo ku salaysan xoojinta gidaarka unugyada inta lagu jiro lignification, subnit ama isku-xirnaanta xeryahooda gaarka ah.Shaqadan waxaa loo aaneyn karaa silico tanjilG_03329 hiddo-wadaha ku dhejinaya anion peroxidase lignin-sameeya oo si weyn loogu habeeyey daraasaddeena 83A: 476 xariiqda u adkaysta ee 6 HPI, laakiin maaha noocyada kale iyo dhibcaha waqti ee aan ka jawaabin.
9S-/13S-lipoxygenase of linoleic acid waa talaabada ugu horeysa ee dariiqa oksaydhka ee biosynthesis ee lipid.Alaabooyinka jidkani waxay leeyihiin hawlo badan oo difaaca dhirta, oo ay ku jiraan xoojinta derbiga unugyada iyada oo loo marayo samaynta kaydka callose iyo pectin, iyo nidaaminta cadaadiska oksaydhka iyada oo loo marayo soo saarista noocyada oksijiinta falcelinta79,80,81,82,83.Daraasadda hadda jirta, muujinta linoleic acid 9S- / 13S-lipoxygenase ayaa lagu beddelay dhammaan noocyada kala duwan, laakiin dadka u nugul 22660, kor u kaca ayaa ka adkaaday waqtiyo kala duwan, halka noocyada kala duwan ee Lanr1 iyo AnMan allele, ay xoogga saarayaan kala-duwanaanta lakabyada oxylipin ee u dhexeeya fal-celinta oxylipin-ka ee geerfahan.
1-aminocyclopropane-1-carboxylate oxidase (ACO) homologue ayaa si weyn kor loogu qaaday (9 hiddo-sideyaasha) ama hoos loo dhigay (2 hiddo-sideyaasha) marka lagu tallaalo lupine.Laba marka laga reebo, dhammaan jawaabahaas waxay ka dhaceen 6 hp.at 83A:476.Dareen-celinta enzymatic ee ay dhexdhexaadiyaan borotiinnada ACO waa heerka xaddidaya heerka wax-soo-saarka ethylene sidaas darteed aad ayaa loo habeeyey84.Ethylene waa hoormoonka dhirta oo ka ciyaara doorar kala duwan nidaaminta horumarinta dhirta iyo ka jawaab celinta abiotic iyo xaaladaha walaaca biotic.Soo saarista qoraalka ACO iyo firfircoonida dariiqa calaamadaynta ethylene waxay ku lug leedahay kordhinta caabbinta bariiska ee fangas hemibiotrophic oryzae oryzae iyadoo nidaaminaysa soosaarka noocyada ogsijiinta falcelinta iyo phytoalexins.Geedi socodka caabuqa caleenta ee la midka ah ee laga helay inta u dhaxaysa M. oryzae iyo C. lupini88,89, oo ka soo horjeeda dib u habeynta muhiimka ah ee ACO homologues ee 83A: 476 ayaa lagu soo warramey daraasaddan, waxay u beddeshaa suurtagalnimada in la keeno caabbinta NLL anthracnose Ethylene oo ah tallaabo dhexe oo calaamad u ah waddooyinka molecular.
Daraasadda hadda jirta, xakamaynta baaxadda weyn ee hiddo-wadaha badan ee la xidhiidha sawir-qaadista ayaa lagu arkay 6 hpi ee 83A: 476 iyo 48 hpi ee Mandeloop iyo 22660 dadweynaha.Baaxadda iyo horumarka isbeddelladani waxay la siman yihiin heerka.Cadaadiska Anthracnose ayaa lagu arkay tijaabadan.Dhawaan, cadaadis xoog leh oo hore oo qoraal ah oo la xidhiidha sawir-qaadista ayaa lagu soo warramey dhowr nooc oo isdhexgalka dhirta-pathogen, oo ay ku jiraan bakteeriyada pathogenic iyo fungi.Degdega (laga bilaabo 2 HPI ee isdhexgalka qaarkood) iyo xakamaynta caalamiga ah ee hiddo-wadaha la xidhiidha photosynthesis ee ka jawaab celinta caabuqa waxay kicin kartaa difaaca dhirta iyadoo lagu saleynayo dejinta noocyada oksijiinta falcelinta iyo isdhexgalka ay la leeyihiin dariiqa salicylic acid si ay u dhexdhexaadiyaan falcelinta xasaasiyadda 90,94.
Gebogebadii, hababka jawaab celinta difaaca ee loo soo jeediyay nasabka ugu adkaysiga badan (83A: 476) waxaa ka mid ah aqoonsiga degdega ah ee pathogen-ka ee hidda-wadaha R (malaha TIR-NBS-LRR TanjilG_05042) iyo jawaabta xasaasiyadeed ee salicylic acid iyo ethylene signaling, oo ay ku xigto sameynta SAR-dheer.ficilka waxaa taageera borotiinka DIR-1.Waa in la ogaadaa in muddada biotrophic ee caabuqa C. lupini uu aad u gaaban yahay (qiyaastii 2 maalmood), oo ay ku xigto koritaanka necrotic95.Kala-guurka u dhexeeya heerarkan waxaa laga yaabaa inay la xiriirto necrosis iyo muujinta borotiinnada ethylene-inducible kuwaas oo u dhaqma sidii kicinta dareen-celinta xasaasiyadeed ee dhirta martida loo yahay.Sidaa darteed, daaqadda waqtiga ee lagu guuleysto qabashada C. lupini ee marxaladda biotrophic waa mid aad u yar.Dib-u-habaynta hiddo-wadaha la xidhiidha redox iyo photosynthesis ee lagu arkay 83A: 476 at 6 hpi waxay la socotaa horumarka fangas hyphae waxayna ku bishaaraynaysaa horumarinta jawaab celin difaac ah oo lagu guuleysto marxaladda biotrophic.Jawaabaha qoraalka ah ee Mandelup iyo dadka 22660 ayaa laga yaabaa inay aad u daahiyaan si ay u qabtaan fungus ka hor intaysan u wareegin koritaanka necrotic, si kastaba ha ahaatee, Mandelup waxaa laga yaabaa inuu ka waxtar badan yahay dadka 22660 sababtoo ah nidaamka degdegga ah ee borotiinka PR-10 wuxuu kor u qaadaa iska caabin toosan.
ETI, oo uu wado hiddo-wadaha R canonical, ayaa u muuqda inuu yahay hab caadi ah oo iska caabbinta digirta ee anthracnose.Sidaa darteed, qaabka legume Medicago truncatula, iska caabbinta anthracnose waxaa bixiya hidda-wadaha RCT1, oo xubin ka ah fasalka TIR-NBS-LRR97 dhirta R.Hiddo-sidahani waxa kale oo uu keena caabbinta anthracnose-ballaaran ee alfalfa marka loo wareejiyo dhirta nugul.Digirta caadiga ah (P. vulgaris), in ka badan laba darsin hiddo-wadaha iska caabinta anthracnose ayaa la aqoonsaday ilaa maanta.Qaar ka mid ah hidde-sideyaashan ayaa laga helaa gobollada ay ka maqan yihiin hiddo-wadaha R-da, si kastaba ha ahaatee qaar kale oo badan ayaa ku yaal cidhifyada koromosoomyada sidda kooxda hidda-wadaha NBS-LRR, oo ay ku jiraan TIR-NBS-LRRs99.Daraasada SSR ee genom-ballaaran ayaa sidoo kale xaqiijisay xiriirka hidda-wadaha NBS-LRR ee leh caabbinta anthracnose ee digirta caadiga ah.Hidde-sidaha R-da ee canonical ayaa sidoo kale laga helay gobolka genomic oo sita meesha ugu weyn ee iska caabbinta anthracnose ee lupine cad 101.
Shaqadeenu waxay muujinaysaa in falcelinta iska caabbinta degdega ah, firfircoonida marxaladda hore ee caabuqa dhirta (doorbidayaa ugu dambayn 12 hpi), waxay si wax ku ool ah uga ilaalisaa lupine caleen-cidhiidhi ah oo ka yimaada anthracnose oo ay keento fungus pathogenic Collelotrichum lupini.Isticmaalka isku xigxiga wax soo saarka sare, waxaanu soo bandhignay muuqaalo kala duwanaansho ah ee hiddo-wadaha iska caabbinta anthracnose ee dhirta NLL oo ay dhexdhexaadiyaan Lanr1 iyo hiddo-wadaha AnMan.Difaaca guusha leh waxa ka mid ah in si taxadir leh loo naqshadeeyo hiddo-wadaha borotiinnada ku lug leh redox, photosynthesis, iyo pathogenesis-ka saacado gudahooda marka geedka uu la kulmo cudur-sidaha.Dareen-celinta ilaalinta la midka ah, laakiin dib-u-dhaca wakhtiga, aad bay waxtar ugu yar yihiin ka ilaalinta dhirta cudurrada.Iska caabbinta kudka ee uu dhexdhexaadiyo hiddo-wadaha Lanr1 waxay u egtahay jawaabta degdegga ah ee caadiga ah ee hidda-wadaha R (difaaca wax-ka-qabashada), halka hidda-wadaha AnMan ay u badan tahay inuu bixiyo jawaab-celin toosan (difaacu ay kiciso qaab-dhismeedka unugyada microbe-ka), taasoo bixisa heer dhexdhexaad ah oo waara.
215 NLL ee loo isticmaalay in lagu baaro calaamadaha anthracnose waxay ka koobnaayeen 74 beerood, 60 xariiq oo lagu helay isgoysyada ama taranka, 5 mutants, iyo 76 jeermisyada duurjoogta ah ama asalka ah.Khadadka ayaa ka kala yimid 17 waddan, gaar ahaan Poland (58), Spain (47), Germany (27), Australia (26), Russia (19), Belarus (7), Italy (5) iyo khadadka kale.oo ka kala yimid 10 wadan.Qalabka waxa kale oo ka mid ah xadadka u adkaysta tixraaca: 83A: 476, Tanjil, Wonga oo sita Lanr1 allele, iyo Mandelup oo sita allele AnMan.Khadadka waxaa laga helay Xogta Kheyraadka Hiddaha ee Yurub Lupine oo ay hayso Poznań Plant Breeding Ltd., Wiatrowo, Poland (Shaxda Dheeraadka ah S1).
Dhirta waxaa lagu koray xaalado la xakameeyey (photoperiod 16 saacadood, heerkulka 25 ° C maalintii iyo 18 ° C habeenkii).Laba nooc oo bayooloji ah ayaa la falanqeeyay.DNA-da waxaa laga soocay caleemaha saddex toddobaad jira iyadoo la adeegsanayo DNeasy Plant Mini Kit (Qiagen, Hilden, Germany) sida uu qabo borotokoolka.Tayada iyo xoojinta DNA-da go'doonsan waxaa lagu qiimeeyay hababka spectrophotometric (NanoDrop 2000; Thermo Fisher Scientific, Waltham, MA, USA).Calaamadaha AnManM1 ee calaamadeynaya hidda-wadaha iska caabbinta anthracnose AnMan (oo laga soo qaatay cv. Mandelup) iyo calaamadeeyayaasha Anseq3 iyo Anseq4 oo garab taagan hidda-wadaha Lanr1 (oo laga soo qaatay cv. Tanjil) ayaa la falanqeeyay 11,26,28.Homozygotes ee allele adkaysi u leh waxa loo dhaliyay sida “1″, u nugul – sida “0”, iyo heterozygotes – sida 0.5.
Iyada oo ku saleysan natiijooyinka baaritaanka calaamadaha AnManM1, AnSeq3 iyo AnSeq4 iyo helitaanka abuurka tijaabooyinka dabagalka ugu dambeeya, 50 khadadka NLL ayaa loo doortay fenotyping caabbinta anthracnose.Falanqaynta ayaa lagu sameeyay nuqul ka mid ah aqalka dhirta lagu koriyo ee kombuyuutarku gacanta ku hayo oo leh 14 saacadood oo sawir ah oo leh heerkul kala duwan oo ah 22 ° C maalintii iyo 19 ° C habeenkii.Abuurka waa la xoqaa (gooynta jaakada abuurka dhinaca ka soo horjeeda ee embriyaha oo leh daab fiiqan) ka hor inta aan la beerin si looga hortago hurdo abuurka sababtoo ah jaakada abuurka oo aad u adag iyo si loo hubiyo biqilka isku midka ah.Dhirta waxaa lagu beeray dheryo (11 × 11 × 21 cm) oo leh carro nadiif ah (TS-1 REC 085 Dhexdhexaad Basic, Klasmann-Deilmann Polska, Warsaw, Poland).Tallaalka waxaa lagu fuliyay Colletotrichum lupini Col-08 cadaadis, koray 1999 ka afkoda dhirta lupine cidhiidhi ah koray beer ku taal Verzhenitsa, Greater Poland (52° 27′ 42″ N 17° 04′ 05″ E).Meel qaadoGo'doominta ayaa lagu dhaqay dhex dhexaadinta XDS oo ah 20° C. iftiinka madow hoostiisa muddo 21 maalmood ah si loo kiciyo xameetida.Afar usbuuc ka dib markii la beero, markii dhirtu ay gaartay heerka caleenta 4-6, tallaalka ayaa lagu sameeyay iyadoo lagu buufiyo xannibaadda conidia ee uruurinta 0.5 x 106 conidia halkii ml.Tallaalka ka dib, dhirta waxaa lagu hayaa mugdi 24 saacadood oo qoyaan ah oo ku saabsan 98% iyo heerkul ah 25 ° C si loo fududeeyo biqilka conidi iyo habka caabuqa.Dhirta ayaa markaas lagu koray 14-saac oo sawir ah oo ah 22°C maalintii/habeenkii 19°C iyo 70% qoyaan.Dhibcaha cudurka ayaa la sameeyay 22 maalmood ka dib tallaalka waxayna u dhaxaysaa 0 (difaaca) ilaa 9 (aad u nugul) iyadoo ku xiran joogitaanka ama maqnaanshaha nabarrada necrotic ee afkoda iyo caleemaha.Intaa waxaa dheer, ka dib dhibcaha, miisaanka dhirta ayaa la qiyaasay.Cilaaqaadka ka dhexeeya genotypes calaamadeeyayaasha iyo noocyada cudurka waxaa loo xisaabiyay sida dhibcaha isku dhafka ah ee laba-xiddigle ah (maqnaanshaha calaamadaha heterozygous ee jaantusyada falanqaynta phenotype caabbinta anthracnose).