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Angustifolius lupine (NLL, Lupinus angustifolius L.) tsire-tsire ne na leguminous da ake amfani dashi don samar da abinci da haɓaka ƙasa.Fadada NLL a duniya a matsayin amfanin gona ya ja hankalin fungi masu kamuwa da cuta da yawa, ciki har da lupine anthracnose, wanda ke haifar da mummunar cutar anthracnose.An yi amfani da allale guda biyu, Lanr1 da AnMan, waɗanda ke ba da ƙarin juriya, a cikin kiwo na NLL, amma har yanzu ba a san hanyoyin ƙwayoyin ƙwayoyin cuta ba.A cikin wannan binciken, an yi amfani da alamun Lanr1 da AnMan don tantance samfuran NLL na Turai.Gwajin maganin alurar riga kafi a cikin yanayi mai sarrafawa ya tabbatar da ingancin duka masu ba da gudummawar juriya.Bambancin Gene mai zurfi shine an yi shi akan wakili mai tsayayya da kuma mai saukin kamuwa.Anthracnose juriya an hade da wuce gona da iri na kwayoyin ontology sharuddan "GO: 0006952 Tsaro Response", "GO: 0055114 Redox Tsari", da "GO: 0015979 Photosynthesis".Bugu da ƙari, layin Lanr1 (83A: 476) ya nuna mahimmancin fassarar fassarar fassarar da sauri bayan ƙaddamarwa, yayin da sauran layin sun nuna jinkirin wannan amsa ta kimanin sa'o'i 42.Amsoshin tsaro suna hade da TIR-NBS, CC-NBS-LRR da kwayoyin NBS-LRR, sunadaran 10 da ke cikin pathogenesis, sunadaran canja wurin lipid, endoglucan-1,3-β-glucosidase, glycine-rich cell cell proteins, da kuma kwayoyin halitta daga hanyar oxygen.Amsoshin farko ga 83A:476, gami da tsantsan danne ƙwayoyin halittar da ke da alaƙa da photosynthesis, sun zo daidai da nasarar kariya a lokacin haɓakar ci gaban ciyayi na ilimin halittar fungi, yana nuna cewa mai tasiri yana haifar da rigakafi.Halin Mandeloop yana raguwa, kamar yadda yake ja a kwance gabaɗaya.
Lupine mai kunkuntar (NLL, Lupinus angustifolius L.) babban hatsi ne mai gina jiki wanda ya samo asali a yankin yammacin Bahar Rum1,2.A halin yanzu ana noman shi azaman amfanin gona ga dabbobi da mutane.Ana kuma la'akari da koren taki a cikin tsarin jujjuyawar amfanin gona saboda gyaran nitrogen ta hanyar daidaita ƙwayoyin ƙwayoyin cuta da haɓaka tsarin ƙasa gabaɗaya.NLL ta yi saurin aiwatar da aikin cikin gida a cikin karnin da ya gabata kuma har yanzu tana cikin matsin lamba mai yawa3,4,5,6,7,8,9,10,11,12.Tare da yaduwar noman NLL, ɗimbin naman gwari na ƙwayoyin cuta sun haɓaka sabbin kayan aikin noma kuma sun haifar da sabbin cututtuka masu lalata amfanin gona. Mafi ban mamaki ga manoman lupine da masu shayarwa shine bayyanar anthracnose, wanda ke haifar da naman gwari, Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Mafi ban mamaki ga manoman lupine da masu shayarwa shine bayyanar anthracnose, wanda ke haifar da naman gwari, Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Наиболее примечательным для фермеров и селекционеров люпина lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Mafi shahara ga manoman lupine da masu shayarwa shine bullar cutar anthracnose da ke haifar da naman gwari na 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，它是由病原真说：最引人注目的是炭疽病的出现，它是由病原真語語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，它是由养者来说，最引人注目的是炭疽病的出现。 Наиболее поразительным для фермеров и селекционеров люпина. Totrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Mafi daukar hankali ga manoman lupine da masu shayarwa shine bullar anthracnose da ke haifar da cututtukan fungus Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13.Rahotannin farko na cutar sun fito ne daga Brazil da Amurka, tare da alamun alamun da suka bayyana a 1912 da 1929 bi da bi.Duk da haka, bayan kimanin shekaru 30, an sanya kwayar cutar a matsayin Colletotrichum gloeosporioids (Penz.) Penz. & Sacc., teleomorph Glomerella cingulata (Stoneman) Spauld. & Sacc., teleomorph Glomerella cingulata (Stoneman) Spauld. & Sacc., телеоморф Glomerella cingulata (Stoneman) Spauld. & Sacc., teleomorph na 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 a cikin Ilimin Halitta. & H. Schrenk,. & H. Schrenk, .da H. Schrenk. & H.施伦克,. & H.施伦克,.da H. Schlenk, .Halin cututtukan farko da aka yi a tsakiyar karni na 20 ya nuna juriya a cikin NLL da rawaya lupine (L. luteus L.) accessions, amma duk farin lupine (L. albus L.) accessions da aka gwada sun kasance masu saukin kamuwa15,16.Nazarin ya nuna cewa ci gaban anthracnose yana hade da haɓakar hazo (zafin iska) da zafin jiki (a cikin kewayon 12-28 ° C), wanda ke haifar da cin zarafi na juriya a yanayin zafi mafi girma17, 18. A gaskiya ma, lokacin da ake buƙata don conidia don germinate da cutar ta fara , ya kasance sau hudu ya fi guntu a 24 ° C (4 hours) fiye da yanayin zafi 1.Don haka, dumamar yanayi da ke ci gaba da haifar da yaduwar cutar anthracnose.Duk da haka, an lura da cutar a Faransa (1982) da Ukraine (1983) a matsayin alamar barazanar da ke tafe, amma masana'antun lupine sun yi watsi da su a lokacin 20,21.Bayan 'yan shekaru, wannan mummunar cuta ta bazu ko'ina cikin duniya kuma ta shafi manyan kasashe masu samar da lupine kamar Australia, Poland da Jamus22,23,24.Bayan barkewar cutar anthracnose a tsakiyar 1990s, babban bincike ya haifar da gano masu ba da taimako da yawa a cikin samfuran NLL19.NLL juriya ga anthracnose ana sarrafa shi ta hanyar manyan alloli guda biyu daban-daban waɗanda aka samo a cikin tushen ƙwayoyin cuta daban-daban: Lanr1 a cikin cultivar Tanjil da Wonga da AnMan a cikin cultivar.Mandalay 25, 26. Waɗannan alleles sun dace da alamomin ƙwayoyin cuta waɗanda ke tallafawa zaɓin ƙwayoyin cuta masu juriya a cikin shirye-shiryen kiwo25,26,27,28,29,30.Layin kiwo mai juriya 83A: 476 yana ɗauke da Lanr1 allele an ketare tare da layin daji mai rauni P27255 don samun yawan RIL da ke keɓancewa don juriya na anthracnose, wanda ya sa ya yiwu a sanya wurin Lanr1 zuwa alamar chromosome NLL-1131, 32, 33. Alignment na taswira tare da juriya na gemu. NLL ya bayyana wurin duk allele uku akan chromosome iri ɗaya (NLL-11), amma a wurare daban-daban29,34,35.Duk da haka, saboda ƙananan adadin RILs da kuma babban nisa na kwayoyin halitta tsakanin alamomi da ma'auni masu dacewa, ba za a iya yanke shawara mai dogara game da kwayoyin halittar su ba.A gefe guda kuma, yin amfani da juzu'in kwayoyin halitta a cikin lupins yana da wahala saboda ƙarancin farfadowar su, wanda ke haifar da magudin ƙwayoyin cuta37.
Ci gaban germplasm na cikin gida yana ɗauke da allele da ake so a cikin jihar homozygous, irin su 83A: 476 (Lanr1) da Mandelup (AnMan), ya buɗe kofa don nazarin juriya na anthracnose a gaban gaban haɗuwa da alleles a cikin al'ummomin daji.Yiwuwar hanyoyin ƙwayoyin ƙwayoyin cuta.Kwatanta martanin tsaro da aka samar ta takamaiman genotypes.Wannan binciken ya kimanta martanin farkon rubutun NLL zuwa rigakafin C. lupini.Da farko, an duba kwamitin germplasm na NLL na Turai mai ɗauke da layukan 215 ta amfani da alamomin kwayoyin halitta waɗanda ke yin alamar Lanr1 da AnMan alleles.Anthracnose phenotyping sannan an yi shi akan layin 50 NLL, wanda aka zaɓa a baya don alamomin ƙwayoyin cuta, ƙarƙashin yanayin sarrafawa.Dangane da waɗannan gwaje-gwajen, layi huɗu da suka bambanta a cikin juriya na anthracnose da Lanr1/AnMan allelic abun da ke ciki an zaɓi su don bambance-bambancen bayanin bayanan kare kariya ta amfani da hanyoyin haɗin gwiwa guda biyu: babban tsarin RNA da ƙididdige PCR na ainihi.
Nuna wani saitin NLL germplasm (N = 215) tare da alamomi Lanr1 (Anseq3 da Anseq4) da AnMan (Anseq4) da AnMan (AnManM1) ya nuna cewa layi ɗaya ne kawai (95726, kusa da Salamanca-b) yana haɓaka “juriya” allele ga duk alamomin 73.5% layi.Layuka goma sha uku sun samar da allunan “mai jurewa” guda biyu na alamar Lanr1, kuma layukan 8 sun samar da “mai jurewa” alleles na Lanr1.alama.Alamar "juriya" na alamar AnMan (Ƙarin Tebura S1).Layi biyu sun kasance heterozygous don alamar Anseq3 da heterozygous ɗaya don alamar AnManM1.Layukan 42 (19.5%) sun ɗauki saɓanin saɓani na Anseq3 da Anseq4 alleles, yana nuna yawan haɗuwa tsakanin waɗannan loci biyu.Anthracnose phenotypes a ƙarƙashin yanayin sarrafawa (Ƙarin Teburin S2) ya bayyana bambance-bambance a cikin juriya na genotypes da aka gwada, wanda aka nuna a cikin tsananin anthracnose.Bambance-bambance a cikin ma'aunin ma'ana sun kasance daga 1.8 (madaidaicin juriya) zuwa 6.9 (mai saukin kamuwa) da bambance-bambancen nauyin shuka ya tashi daga 0.62 (mai saukin kamuwa) zuwa 4.45 g (mai jurewa). Akwai mahimmiyar alaƙa tsakanin ƙimar da aka lura a cikin maimaitawa guda biyu na gwajin (0.51 don ƙimar cututtukan cuta, P = 0.00017 da 0.61 don nauyin shuka, P <0.0001) da kuma tsakanin waɗannan sigogi biyu (- 0.59 da - 0.77, P <0.0001). Akwai mahimmiyar alaƙa tsakanin ƙimar da aka lura a cikin nau'ikan gwaji guda biyu (0.51 don ƙimar cututtukan cuta, P = 0.00017 da 0.61 don nauyin shuka, P <0.0001) da kuma tsakanin waɗannan sigogi biyu (- 0.59 da - 0.77, P <0.0001). Выявлена ​​достоверная корреляция межdu zanachенияmi, наблюdaemыmy в двух повторностях эksperyatsyonnaya , 0,51 . 0,00017 da 0,61 da sauransu, P <0,0001), da sauransu. An sami muhimmiyar alaƙa tsakanin ƙimar da aka lura a cikin maimaitawar gwaji guda biyu (0.51 don ƙimar cutar, P = 0.00017 da 0.61 don nauyin shuka, P <0.0001), da kuma tsakanin waɗannan sigogi biyu (- 0.59 da -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 ， p <0.0001）和 - 0.77, P <0.0001). Наблюдалась значительная корреляция между значениями. 017 da kuma massа растения 0,61, P <0,0001), da dai sauransu Akwai mahimmin alaƙa tsakanin ƙimar da aka lura a cikin kwafin (ƙimar rashin lafiya 0.51, P = 0.00017 da nauyin shuka 0.61, P <0.0001) da tsakanin waɗannan sigogi guda biyu (-0.59 da -0 .0001) 0.77, P <0.0001. ).Alamun alamomin da ake gani a cikin tsire-tsire masu saurin kamuwa sun haɗa da ƙwanƙwasa da karkatar da tushe mai kama da tsarin “bakan makiyayi”, sannan kuma raunukan oval tare da sporozoites orange/ ruwan hoda (Ƙarin Hoto 1).Hanyoyin shiga Ostiraliya dauke da Lanr1 (83A: 476 da Tanjil) da kuma kwayoyin AnMan (Mandelup) suna da matsakaicin juriya, 0.0331 da 0.0036).Wasu layukan da ke ɗauke da “mai jurewa” Lanr1 da/ko AnMan alleles suna nuna alamun cutar.
Abin sha'awa shine, 'yan layin NLL da ba su da duk wani alamar "mai jurewa" alama sun nuna babban matakin juriya na anthracnose (kwatankwacin ko mafi girma fiye da Lanr1 ko AnMan genotypes), irin su Boregine (P darajar <0.0001 ga duka sigogi), Bojar (P darajar <0.0001 don ci da 0.0001 Population ga shuka-5) 4 da kuma 0.001 don nauyin shuka-5. kuma maras muhimmanci ga nauyi). Abin sha'awa shine, 'yan layin NLL da ba su da duk wani alamar "mai jurewa" alama sun nuna babban matakin juriya na anthracnose (kwatankwacin ko mafi girma fiye da Lanr1 ko AnMan genotypes), irin su Boregine (P darajar <0.0001 ga duka sigogi), Bojar (P darajar <0.0001 don ci da 0.0001 Population ga shuka-5) 4 da kuma 0.001 don nauyin shuka-5. kuma maras muhimmanci ga nauyi). Интересно, что несколько линий NLL, лишенных какого-лиBO ости к антракнозу ). ). Abin sha'awa, yawancin layin NLL da ba su da duk wani alamar alamar 'resistant' sun nuna babban matakin juriya ga anthracnose (kwatankwacin ko mafi girma fiye da Lanr1 ko AnMan genotypes), irin su Boregine (P darajar <0.0001 ga duka sigogi), Bojar (P darajar <0.0001 don kimantawa da kuma 0.0001) yawan jama'a B. 1 don kimantawa kuma ba mahimmanci ga nauyi ba).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） B-549/79b（得分P 值< 0.0001，重量不显着）。 Yana da ban sha'awa cewa wasu tsarin NLL waɗanda ba su da alamun "antigenic" suna nuna juriya mai tsayi (daidai da Lanr1 ko AnMan genes ko mafi girma), irin su Boregine (duka sigogi P <0.0001), Bojar ( darajar P <0.0001, nauyin shuka 0.001) da kuma nau'in B-549 / 0.0 . Интересно, что некоторые линии NLL, NLL к антракнозу (сравнимые или выше, чем у генотипов Lanr1 или AnMan), такие как Boregine (значение P для обоих повыш0, 000), Boregine 1, масса растения 0,001) da kuma популяция B-549/79b (оценка P-значение <0,0001, масса незначительна). Abin sha'awa, wasu layin NLL da ba su da duk wani alamar alamar 'juriya' sun nuna babban matakan juriya na anthracnose (kwatankwacin ko mafi girma fiye da Lanr1 ko AnMan genotypes), irin su Boregine (ƙimar P don duka sigogi <0.0001) , Bojar (P-darajar <0.0001, nauyin shuka 0.00-051) da kuma yawan jama'a 0.0005 (P) nauyi ba shi da mahimmanci).Wannan sabon abu yana ba da shawarar yiwuwar wani sabon tushen juriya na kwayoyin halitta, yana bayyana rashin daidaituwa tsakanin alamomin genotypes da cututtukan cututtukan cututtuka (ƙimar P daga ~ 0.42 zuwa ~ 0.98).Don haka, gwajin Kolmogorov-Smirnov ya nuna cewa an rarraba bayanan akan juriya na anthracnose kusan yawanci don maki (P-darajar 0.25 da 0.11) da kuma yawan shuka (P-darajar 0.47 da 0.55), yana ba da shawarar cewa ƙarin allele fiye da Lanr1 da AnMan suna da hannu.
Dangane da sakamakon gwajin juriya na anthracnose, an zaɓi layukan 4 don nazarin fassarar: 83A: 476, Boregine, Mandelup, da Yawan jama'a 22660. An sake gwada waɗannan layin don juriya na anthrax a cikin gwaje-gwajen inoculation ta hanyar RNA, muddin sun kasance daidai da na gwajin baya.Mahimman ƙimar sun kasance kamar haka: Boregin (1.71 ± 1.39), 83A: 476 (2.09 ± 1.38), Mandelup (3.82 ± 1.42) da yawan jama'a 22660 (6.11 ± 1.29).
Yarjejeniyar Illumina NovaSeq 6000 ta sami matsakaicin nau'i-nau'i na Mread 40.5 a kowane samfurin (29.7 zuwa 54.4 Mreads) (Ƙarin Teburin S3).Makiyoyin daidaitawa a cikin jeri na tunani sun kasance daga 75.5% zuwa 88.6%.Matsakaicin daidaitawar bayanan kirgawa tsakanin bambance-bambancen gwaji tsakanin kwafi na halitta ya tashi daga 0.812 zuwa 0.997 (ma'ana 0.959). Daga cikin kwayoyin halittar 35,170 da aka bincika, 2917 bai bayyana ba, kuma sauran nau'ikan 4785 an bayyana su a matakin da ba su da kyau (tushe yana nufin <5). Daga cikin kwayoyin halittar 35,170 da aka bincika, 2917 bai bayyana ba, kuma sauran nau'ikan 4785 an bayyana su a matakin da ba su da kyau (tushe yana nufin <5). Иz 35 170 proanalizirovannыh genova 2917 овне (bazовое среднее <5). Daga cikin kwayoyin 35,170 da aka bincikar, 2917 ba ta nuna wani magana ba, kuma sauran kwayoyin 4785 an bayyana su a matakin da ba shi da kyau (tushe yana nufin <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 da kuma m 4785 е среднее значение <5). Daga cikin kwayoyin halitta 35,170 da aka bincika, 2917 ba a bayyana ba kuma sauran kwayoyin 4785 suna da furci mara kyau (tushe yana nufin <5).Don haka, adadin kwayoyin halittar da aka yi la'akari da su (tushe yana nufin ≥ 5) yayin gwajin shine 27,468 (78.1%) (Ƙarin Teburin S4).
Daga farkon lokaci, duk layin NLL sun amsa maganin C. lupini (rashin Col-08) ta hanyar sake tsara rubutun (Table 1), duk da haka, an sami bambance-bambance masu mahimmanci tsakanin layin.Don haka, layin juriya na 83A: 476 (dauke da Lanr1 gene) ya nuna gagarumin reprogramming reprogramming a farkon lokaci (6 hpi) tare da karuwar 31-69-ninki a cikin adadin keɓaɓɓen sama-da ƙasa-genes idan aka kwatanta da sauran lokutan lokaci a wannan lokacin.Bugu da kari, wannan kololuwar ba ta dadewa ba, yayin da furucin 'yan tsirarun kwayoyin halitta ya ragu sosai a karo na biyu (12 hpi).Abin sha'awa shine, Boregine, wanda kuma ya nuna juriya mai yawa a cikin gwajin gwaji, ba a yi irin wannan babban tsarin sake fasalin ba yayin gwajin.Koyaya, adadin jinsin da aka bayyana daban-daban (DEG) iri ɗaya ne ga Boregine da 83A: 476 a 12 HPI.Dukansu Mandelup da yawan jama'a 22660 sun nuna ƙimar DEG a ƙarshen lokaci (48 l / s), yana nuna jinkirin dangi a cikin martanin tsaro.
Saboda 83A: 476 an yi reprogramming mai girma a mayar da martani ga C. lupini a 6 HPI idan aka kwatanta da duk sauran layi, ~ 91% na DEGs da aka lura a wannan lokacin sun kasance takamaiman jinsi (Fig. 1).Duk da haka, akwai wasu rikice-rikice a farkon martani tsakanin layin binciken, kamar yadda 68.5%, 50.9%, da 52.6% DEG a cikin Boregine, Mandelup, da yawan jama'a 22660, bi da bi, tare da waɗanda aka samu a 83A: 476 a wasu lokuta a cikin lokaci.Koyaya, waɗannan DEGs sun ƙididdige ɗan ƙaramin juzu'i (0.97-1.70%) na duk DEGs a halin yanzu da aka gano ta amfani da 83A: 476.Bugu da ƙari, 11 DEGs daga duk layi sun kasance masu daidaituwa a wannan lokacin (Ƙarin Tables S4-S6), ciki har da abubuwan gama gari na amsawar kariya ta shuka: furotin canja wurin lipid (TanjilG_32225), endoglucan-1,3-β-glucoside enzyme (TanjilG_23384), sunadarai guda biyu na damuwa-inducible kamar SAM2G23 (TanjilG3) ​​da Tanjil33 (15G_5). furotin tex (TanjilG_32352), da kuma sunadaran bangon bango mai wadatar glycine guda biyu (TanjilG_19701 da TanjilG_19702).Har ila yau, an sami babban madaidaici a cikin martanin da aka rubuta tsakanin 83A: 476 da Boregine a 24 HPI (jimlar 16-38% DEG) da tsakanin Mandelup da Yawan jama'a 22660 a 48 HPI (jimlar 14-20% DEG).
Zane na Venn yana nuna adadin nau'ikan kwayoyin halitta daban-daban (DEG) a cikin layin lupine (NLL) kunkuntar da aka yi tare da Colletotrichum lupini (danniya Col-08 da aka samu daga filayen lupine a Wierzhenice, Poland, 1999).Layukan NLL da aka bincika sune: 83A: 476 (mai jurewa, ɗauke da Lanr1 allele), Boregine (mai jurewa, asalin halitta wanda ba a san shi ba), Mandelup (mai juriya mai matsakaici, ɗauke da AnMan allele) da yawan jama'a 22660 (mai saukin kamuwa).Gajartawar hpi tana tsaye na sa'o'i bayan alurar riga kafi.An cire ƙimar sifili don sauƙaƙe jadawali.
An yi nazari akan saitin kwayoyin halittar da aka wuce gona da iri a 6 hpi don kasancewar sassan halittu na canonical R (Ƙarin Teburin S7).Wannan binciken ya bayyana ƙaddamar da ƙayyadaddun ƙayyadaddun kwayoyin halittar juriya na cututtuka tare da yankunan NBS-LRR kawai a 83A: 476.Wannan saitin ya ƙunshi nau'in TIR-NBS-LRR guda ɗaya (tanjilg_05042), ƙwayoyin CC-NBS-LRR guda biyar (tanjilg_06165, tanjilg_06162, tanjilg_22773, tanjilg_22640, da tanjilg_1616-LR_4), da huɗu N1 N1BSILR_2 (tanjilg_16162) da kuma NBS-Lrr hudu (tanjilg_16162) da NBS-LRR hudu (TANJILG_16162).Duk waɗannan kwayoyin halitta suna da yanki na canonical da aka tsara a jeri-jeri.Baya ga kwayoyin halittar NBS-LRR, an kunna kinases RLL da yawa a 6 hpi, wato daya a cikin Boregine (TanjilG_19877), biyu a cikin Mandelup (TanjilG_07141 da TanjilG_19877) kuma a cikin yawan jama'a 22660 (TanjilG_0901) .
Kwayoyin halitta tare da mahimmancin magana mai mahimmanci don mayar da martani ga allurar rigakafi tare da C. lupini (rashin Col-08) an ƙaddamar da bincike na haɓakawa na Gene Ontology (GO) (Ƙarin Table S8). Mafi yawan lokuta da aka fi dacewa da tsarin tsarin nazarin halittu shine "GO: 0006952 amsawar tsaro" wanda ya bayyana a cikin 6 daga cikin 16 (lokaci × layi) haɗuwa tare da babban mahimmanci (P darajar <0.001) (Fig. 2). Mafi yawan lokuta da aka fi dacewa da tsarin tsarin nazarin halittu shine "GO: 0006952 amsawar tsaro" wanda ya bayyana a cikin 6 daga cikin 16 (lokaci × layi) haɗuwa tare da babban mahimmanci (P darajar <0.001) (Fig. 2). Наиболеечасто чрезмерно представленным термином я 6 da 16 Mafi yawan lokuta da aka fi nunawa akan tsarin tsarin halitta shine 'GO: 0006952 amsawar tsaro', wanda ya bayyana a cikin 6 na 16 (lokaci × layi) haɗuwa tare da babban mahimmanci (P darajar <0.001) (Fig. 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）。 Mafi yawan wakilcin tsarin tsarin halitta shine "GO: 0006952 amsawar tsaro", wanda ya bayyana a cikin 6 na 16 (时间×线) haɗuwa, tare da babban mahimmanci (P darajar <0.001) (图2) . Наиболее часто чрезмерно представленным термином биологического процесса был «GO: 0006952 Amsar Tsaro», 1 комбинаций (время × линия) с высокой значимостью (значение P <0,001) (рис. 2). Mafi yawan lokuta da aka fi yawan wakilcin tsarin tsarin halitta shine 'GO: 0006952 Response Defence', wanda ya bayyana a cikin 6 daga cikin 16 haɗuwa (layi × lokaci) tare da mahimmanci (P darajar <0.001) (Fig. 2).An ba da wannan kalmar a lokaci biyu a cikin 83A: 476 da Boregine (6 da 24 hpi) kuma a lokaci ɗaya a cikin Mandelup da Yawan jama'a 22660 (12 da 6 hpi, bi da bi).Wannan shine sakamakon da ake tsammanin, yana nuna alamar maganin antifungal na layukan masu juriya.Bugu da ƙari, 83A: 476 ya amsa ga C. lupini ta hanyar saurin haifar da kwayoyin halitta da ke da alaka da fashewar oxidative da aka wakilta ta kalmar "GO: 0055114 redox", yana nuna wani takamaiman martani na tsaro, yayin da Boregine ya bayyana takamaiman martani na tsaro, dangane da kalmar 'GO'.:0006950 Amsar damuwa”.Yawan jama'a 22660 ya kunna amsawar juriya a kwance wanda ya shafi metabolites na biyu, yana nuna yawan adadin sharuɗɗan "GO: 0016104 Tsarin biosynthesis na triterpene" da "GO: 0006722 Tsarin triterpene metabolism" (dukkanin sharuɗɗan sun kasance cikin saitin kwayoyin halitta guda ɗaya), la'akari da sakamakon kwanciyar hankali, tsakanin ƙarin haɓakawa da haɓakawa 6. , Farkon amsawa 83A: 476 (6 hpi) da jinkirin amsawar Mandelup da Yawan jama'a 22660 sun haɗa da kalmar GO: 0015979 'photosynthesis' da sauran hanyoyin nazarin halittu.
Sharuɗɗan kwayoyin halitta na ontology da aka zaɓa a cikin bayanin kwayoyin halittar da aka bayyana daban-daban yayin amsawar lupine mai kunkuntar (NLL) da aka yi da lupine anthrax (irin Col-08 da aka samu daga filayen lupine a Wierzhenice, Poland, a cikin 1999) an wuce gona da iri sosai.Layukan NLL da aka bincika sune: 83A: 476 (mai jurewa, ɗauke da homozygous Lanr1 allele), Boregine (mai juriya, asalin halittar da ba a sani ba), Mandelup (mai juriya mai matsakaici, ɗauke da homozygous AnMan allele) da yawan jama'a 22660 (mai yiwuwa).
Saboda wannan binciken ya yi niyya don gano kwayoyin halittar da ke ba da gudummawa ga juriya na anthracnose, kwayoyin da aka sanya wa sharuɗɗan GO "GO: 0006952 Amsoshin Tsaro" da "GO: 0055114 Redox tafiyar matakai" an yi nazari tare da yanke-offs tun asali yana nufin ≥ 30 tare da akalla layi daya.× aya a cikin lokacin haɗa mahimman ƙimar log2 (canji ninki).Adadin kwayoyin halittar da suka cika wadannan sharudda shine 65 na GO:0006952 da 524 don GO:0055114.
83A: 476 ya bayyana kololuwar DEG guda biyu da aka bayyana tare da kalmar GO: 0006952, na farko a cikin kwayoyin halitta 6 a kowace inch (jiniyoyin 64, ka'idojin sama da ƙasa) da na biyu a cikin ƙwayoyin 24 a kowace inch (jinin 15, ƙa'ida kawai).Boregine kuma ya nuna cewa GO: 0006952 ya yi girma a lokaci guda, amma tare da ƙarancin DEG (11 da 8) da kunna fifiko.Mandeloop ya nuna kololuwa biyu na GO: 0006952 a 12 da 48 HPI, dukansu suna ɗauke da kwayoyin halitta 12 (na farko tare da kwayoyin halitta masu kunnawa, kuma na biyu tare da ƙwayoyin cuta kawai), yayin da yawan mutanen 22660 a 6 HPI (13 genes) suna da fifiko mafi girma na ƙãra kololuwa.tsari.Ya kamata a lura cewa 96.4% na GO: 0006952 DEG a cikin waɗannan kololuwa suna da nau'in amsa iri ɗaya (sama ko ƙasa), yana nuna mahimmancin haɗuwa a cikin martanin tsaro duk da bambance-bambance a cikin adadin kwayoyin halitta.Mafi girman rukuni na jerin abubuwan da ke da alaƙa da kalmar GO: 0006952 tana ɓoye Protein Protein 22 (SAM22-kamar), wanda ke cikin nau'in furotin mai alaƙa da pathogenesis (PR-10) da core protein latex.irin (MLP-like) furotin) furotin (Fig. 3).Ƙungiyoyin biyu sun bambanta ta yanayin magana da kuma alkiblar mayar da martani.Kwayoyin halittar da ke ɓoye sunadaran SAM22-kamar sun nuna daidaito da mahimmancin shigarwa a farkon lokacin (6 ko 12 hpi) kuma gabaɗaya ba su da amsa a ƙarshen gwajin (48 hpi), yayin da sunadarai masu kama da MLP sun nuna daidaituwa a 6 hpi.hpi.83A: 476 da Mandelup a 48 hp/in, kusan duk sauran bayanan ba su da amsa.Bugu da ƙari, bambance-bambance a cikin bayanan bayanan furotin na SAM22-kamar sunadaran sunadaran sun biyo bayan lura da sauye-sauye a cikin juriya na anthracnose, yayin da mafi yawan layin da ke da tsayin daka yana da mafi yawan lokaci da ke haifar da waɗannan kwayoyin fiye da kwayoyin halitta masu saukin kamuwa.Wani nau'in LlR18A/B-kamar PR-10 ya nuna nau'in magana mai kama da kwayar halitta kamar SAM22.
An gano manyan abubuwan da ke cikin tsarin tsarin ilimin halitta "GO: 0006952 Amsar Tsaro" da kuma tsarin maganganun 'yan takara na Lanr1 da AnMan alleles.Ma'auni na Log2 yana wakiltar ƙimar log2 (canjin ninka) tsakanin inoculated (Colletotrichum lupini, damuwa Col-08, wanda aka samo daga filayen lupine, Wizhenica, Poland, 1999) da sarrafawa (sham-inocated) tsire-tsire a lokaci guda.An yi nazarin layukan lupine narrowleaf masu zuwa: 83A: 476 (mai jurewa, ɗauke da homozygous Lanr1 allele), Boregine (mai jurewa, asalin halitta wanda ba a san shi ba), Mandelup (mai juriya mai matsakaici, ɗauke da homozygous AnMan allele), da yawan jama'a 22660 (mai yiwuwa).
Bugu da ƙari, an kimanta bayanan bayanan RNA-seq ɗan takarar ɗan takarar Lanr1 (TanjilG_05042) da AnMan (TanjilG_12861) (Fig. 3).Halin TanjilG_05042 ya nuna muhimmiyar amsa (kunnawa) a 83A: 476 kawai a farkon lokacin (6 hpi), yayin da TanjilG_12861 ya kasance mai mahimmanci a cikin Mandeloop kawai a maki biyu: 6 hpi (ka'idar ƙasa) da 24 hpi (6 hpi).Da.).daidaitacce)).
Mafi yawan kwayoyin halitta a cikin kalmar GO: 0055114 "tsarin redox" sune kwayoyin da ke ɓoye sunadaran cytochrome P450 da peroxidase (Fig. 4).Don samfuran da aka keɓe daga 83A: 476 a 6 HPI, matsakaicin ko mafi ƙarancin log2 (canjin ninka) ƙimar (na 86.6% na kwayoyin halitta) ana lura da su gabaɗaya tsakanin tsire-tsire masu inoculated da sarrafawa, yana nuna babban martani na wannan genotype zuwa inoculating jima'i.83A: 476 ya nuna GO mafi mahimmanci: 0055114 DEG a 6 hpi (503 genes), yayin da sauran layin a 48 hpi (Boregine, 31 genes; Mandelup, 85 genes; da Population 22660, 78 genes)).A cikin mafi yawan kwayoyin halittar GO: 0055114 iyali, an lura da nau'i biyu na martani ga allurar rigakafi (kunnawa da hanawa).Abin sha'awa, har zuwa 97.6% na DEGs da aka gano don kalmar GO: 0055114 a cikin Mandelupe a 48 hp Wadannan abubuwan lura sun nuna cewa, duk da ƙananan ƙananan sikelin (watau adadin kwayoyin halittar redox da aka canza, 85 da 503), tsarin jinkirin martanin da aka ba da amsa na manracnose na 6 shine farkon 8.3.A cikin Boregine da Yawan jama'a 22660, wannan haɗuwa ya yi ƙasa da 51.6% da 75.6%, bi da bi.
An bayyana alamun bayyanar manyan abubuwan da ke cikin kalmar tsarin nazarin halittu "GO: 0055114 Redox".Ma'auni na Log2 yana wakiltar ƙimar log2 (canjin ninka) tsakanin inoculated (Colletotrichum lupini, damuwa Col-08, wanda aka samo daga filayen lupine, Wizhenica, Poland, 1999) da sarrafawa (sham-inocated) tsire-tsire a lokaci guda.An yi nazarin layukan lupine narrowleaf masu zuwa: 83A: 476 (mai jurewa, ɗauke da homozygous Lanr1 allele), Boregine (mai jurewa, asalin halitta wanda ba a san shi ba), Mandelup (mai juriya mai matsakaici, ɗauke da homozygous AnMan allele), da yawan jama'a 22660 (mai yiwuwa).
83A: 476 Amsoshi na fassarar fassarar zuwa inoculation tare da C. lupini (irin Col-08) kuma ya haɗa da haɗin gwiwar shiru na kwayoyin halitta wanda aka danganta ga kalmar GO: 0015979 "photosynthesis" da sauran hanyoyin nazarin halittu masu dangantaka (FIG. 5).Wannan GO:0015979 DEG saitin ya ƙunshi kwayoyin halitta 105 waɗanda aka danne sosai a 6 hpi a 83A:476.A cikin wannan juzu'in, an kuma lalata kwayoyin halittar 37 a cikin Mandelup a 48 HPI da 35 a lokaci guda a cikin yawan 22660, gami da 19 DEGs gama gari ga duka genotypes.Babu DEGs masu alaƙa da kalmar GO: 0015979 da aka kunna sosai a cikin kowane haɗuwa (lokaci x layi).
An bayyana alamun bayyanar manyan abubuwan da ke cikin kalmar tsarin nazarin halittu "GO: 0015979 Photosynthesis".Ma'auni na Log2 yana wakiltar ƙimar log2 (canjin ninka) tsakanin inoculated (Colletotrichum lupini, damuwa Col-08, wanda aka samo daga filayen lupine, Wizhenica, Poland, 1999) da sarrafawa (sham-inocated) tsire-tsire a lokaci guda.An yi nazarin layukan lupine narrowleaf masu zuwa: 83A: 476 (mai jurewa, ɗauke da homozygous Lanr1 allele), Boregine (mai jurewa, asalin halitta wanda ba a san shi ba), Mandelup (mai juriya mai matsakaici, ɗauke da homozygous AnMan allele), da yawan jama'a 22660 (mai yiwuwa).
Dangane da sakamakon bincike na bambance-bambancen magana da kuma yiwuwar shiga cikin martanin tsaro game da fungi na pathogenic, an zaɓi wannan saitin kwayoyin halitta guda bakwai don ƙididdige bayanan bayanan magana ta ainihin lokaci PCR (Ƙarin Table S9).
Halin furotin da aka sakawa TanjilG_10657 an jawo shi sosai a cikin duk layin da aka yi nazari da maki lokaci idan aka kwatanta da sarrafa (mimic) shuke-shuke (Ƙarin Tables S10, S11).Bugu da ƙari, bayanin martaba na TanjilG_10657 ya nuna haɓakar haɓakawa a tsawon lokacin gwajin ga dukkan layi.Yawan jama'a 22660 ya nuna mafi girman hankali na TanjilG_10657 zuwa inoculation tare da kunnawa sau 114 da kuma matakin magana mafi girma (4.4 ± 0.4) a 24 HPI (Fig. 6a).Tsarin furotin na PR10 LlR18A TanjilG_27015 kuma ya nuna kunnawa a duk layin da maki lokaci, tare da mahimmancin ƙididdiga a mafi yawan bayanan bayanai (Fig. 6b).Mai kama da TanjilG_10657, an lura da matakin magana mafi girma na TanjilG_27015 a cikin 22660 da aka yi wa allurar a 24 HPI (19.5 ± 2.4).Acid endochitinase gene TanjilG_04706 an inganta shi sosai a cikin dukkan layi kuma a kowane lokaci ban da Boregine 6 hpi (Fig. 6c).An jawo shi da ƙarfi a farkon lokacin (6 HPI) a 83A: 476 (ta sau 10.5) kuma an ƙara matsakaici a cikin wasu layin (ta sau 6.6-7.5).A lokacin gwajin, bayanin TanjilG_04706 ya kasance a irin wannan matakan a cikin 83A: 476 da Boregine, yayin da a cikin Mandelup da Population 22660 ya karu sosai, ya kai ingantattun ƙima (5.9 ± 1.5 da 6.2 ± 1.5, bi da bi).endoglucan-1,3-β-glucosidase-like gene TanjilG_23384 ya nuna babban kunnawa a farkon lokutan biyu na farko (6 da 12 hpi) a cikin dukkanin layi banda yawan 22660 (Fig. 6d).An lura da matakan maganganun dangi mafi girma na TanjilG_23384 a karo na biyu (12 hpi) a cikin Mandelup (2.7 ± 0.3) da 83A: 476 (1.5 ± 0.1).A 24 HPI, TanjilG_23384 magana ya kasance kaɗan kaɗan a duk layin da aka yi nazari (daga 0.04 ± 0.009 zuwa 0.44 ± 0.12).
Bayanan bayanan bayanan da aka zaɓa (ag) da aka bayyana ta PCR mai ƙididdigewa.Lambobin 6, 12 da 24 suna wakiltar sa'o'i bayan rigakafin.An yi amfani da kwayoyin halittar LanDExH7 da LanTUB6 don daidaitawa kuma an yi amfani da LanTUB6 don daidaita tsakanin jeri.Sandunan kurakurai suna wakiltar daidaitaccen karkatacciyar hanya dangane da kwafi na halitta guda uku, kowanne ɗayansu shine matsakaicin kwafin fasaha uku. Mahimman ƙididdiga na bambance-bambance a cikin matakan magana tsakanin inoculated (Colletotrichum lupini, damuwa Col-08, wanda aka samu a cikin 1999 daga filin lupine a Wierzenica, Poland) da kuma sarrafawa (ba'a-inoculation) tsire-tsire suna alama a sama da bayanan bayanai (* darajar P <0.05, ** P darajar ≤ 0.01 , 1). Mahimman ƙididdiga na bambance-bambance a cikin matakan magana tsakanin inoculated (Colletotrichum lupini, damuwa Col-08, wanda aka samu a cikin 1999 daga filin lupine a Wierzenica, Poland) da kuma sarrafawa (ba'a-inoculation) tsire-tsire suna alama a sama da bayanan bayanai (* darajar P <0.05, ** P darajar ≤ 0.01 , 1). Сатическая значимость различий в уровнях эkspressy межdu инкулированыmy (Colletotrichum lupini, shtamm Col-08, 99, люпина в Верженице, Польша) и контрольными P ≤ 0,01, *** значение P ≤ 0,001). Ƙididdigar bambance-bambance a cikin matakan magana tsakanin inoculated (Colletotrichum lupini, damuwa Col-08, wanda aka samu a cikin 1999 daga filin lupine a Wierzhenice, Poland) da kuma sarrafawa (sham-inocated) tsire-tsire ana lura da su a sama da bayanan bayanai (* darajar P <0.05, ** P-darajar ≤ 0.01, *** - 1).接种（Colletotrichum lupini，Col-08株，1999年从波兰Wierzenica的羽扇豆田获得）和对照（模拟接照（模拟接科又模拟接科剤來來來）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 的 羽扇 获得） 和 对照差异 的 统计学 显着性 标记 数据点 上方*p 值 <0.05, **P ≤ 0.01, ***P ≤ 0.001)). Сатически значимыe razlychyya v urovnyah эkspressyy mezhdu inokulyrovannymy Верженице, Польша, в 1999 г.) и контрольными -значение ≤ 0,01, *** P-значение ≤ 0,001). Ƙididdigar bambance-bambance a cikin matakan magana tsakanin inoculated (Colletotrichum lupini, damuwa Col-08, wanda aka samo daga filayen lupine a Verzhenice, Poland, a cikin 1999) da kuma sarrafawa (sham-inocated) tsire-tsire ana lura da su a sama da bayanan bayanai (* darajar P <0.05, ** P-darajar ≤ 0.01, *** P-darajar ≤ 0.01, *** P)Layukan NLL da aka bincika sune: 83A: 476 (mai jurewa, ɗauke da homozygous Lanr1 allele), Mandelup (mai juriya mai matsakaici, ɗauke da homozygous AnMan allele), Boregine (mai juriya, asalin halittar da ba a sani ba) da yawan jama'a 22660 (mai yiwuwa).
Halin ɗan takara TanjilG_05042 a wurin Lanr1 ya nuna salon magana daban-daban daga bayanan martaba da aka samu daga nazarin RNA-seq (Fig. 6e).An lura da mahimmancin kunna wannan kwayar halitta a cikin Mandelup da yawan 22660 (har zuwa 39.7 da 11.7 sau, bi da bi), wanda ya haifar da ƙananan matakan magana (har zuwa 1.4 ± 0.14 da 7.2 ± 1.3, bi da bi).83A: 476 kuma ya bayyana wasu haɓakawa na TanjilG_05042 gene (har zuwa 3.8-ninka), duk da haka, matakan maganganun dangi da aka samu (0.044 ± 0.002) sun kasance fiye da 30-ninka fiye da waɗanda aka gani a Mandelup da 22660 yawan jama'a.An bincika ta qPCR ya nuna bambance-bambance masu mahimmanci a cikin matakan magana tsakanin genotypes a cikin bambance-bambancen izgili-alurar rigakafi (control), yana kaiwa ga bambancin 58-ninki tsakanin yawan jama'a 22660 da 83A: 476, da kuma tsakanin yawan jama'a 22660 da 22660. An sami bambanci sau biyu tsakanin Boregine da Mandalup.
Halin ɗan takara a wurin AnMan, TanjilG_12861, an kunna shi don amsa allurar rigakafi a cikin 83A: 476 da Mandelup, ya kasance tsaka tsaki a cikin yawan 22660, kuma an rage shi a cikin Boregine (Fig. 6f).Maganar dangi na kwayar halittar TanjilG_12861 ita ce mafi girma a cikin allurar 83A: 476 (0.14± 0.01).Ajin 17.4 kDa I heat shock protein gene TanjilG_05080 HSP17.4 ya nuna ƙananan matakan magana a cikin duk nau'ikan da aka yi nazari da maki lokaci (Fig. 6g).An lura da ƙimar mafi girma a 24 HPI a cikin yawan 22660 (0.14 ± 0.02, karuwa sau takwas don amsa maganin alurar riga kafi).
Kwatanta bayanan bayanan kwayoyin halitta (Fig. 7) ya nuna babban dangantaka tsakanin TanjilG_10657 da wasu kwayoyin halitta guda hudu: TanjilG_27015 (r = 0.89), TanjilG_05080 (r = 0.85), TanjilG_05042 (r = 0.80) (r = 0.80), da Tanjil.Irin waɗannan sakamakon na iya nuna haɗin gwiwar waɗannan kwayoyin halitta yayin martanin tsaro.Kwayoyin TanjilG_12861 da TanjilG_23384 sun nuna bayanan martaba daban-daban tare da ƙananan ƙimar daidaituwar Pearson (daga 0.08 zuwa 0.43 da -0.19 zuwa 0.28, bi da bi) idan aka kwatanta da sauran kwayoyin halitta.
An gano alaƙa tsakanin bayanan bayanan kwayoyin halitta ta amfani da PCR mai ƙididdigewa.An yi nazarin layukan lupine narrowleaf masu zuwa: 83A: 476 (mai jurewa, ɗauke da homozygous Lanr1 allele), Mandelup (mai juriya mai matsakaici, yana ɗauke da homozygous AnMan allele), Boregine (mai jurewa, asalin kwayoyin halitta ba a sani ba), da yawan jama'a 22660 (mai yiwuwa).An ƙididdige maki uku na lokaci (6, 12 da 24 hours bayan inoculation), ciki har da inoculated (Colletotrichum lupini, strain Col-08, samu daga lupine filayen a Wierzhenice, Poland, a 1999) da kuma sarrafawa (sham-inoculation).Ma'auni yana nuna ƙimar haɗin haɗin gwiwar Pearson.
Dangane da bayanan da aka samu a 6 horsepower a kowace inch, WGCNA da aka yi a kan 9981 DEG gano ta hanyar kwatanta inoculated da sarrafawa shuke-shuke don mayar da hankali a kan farkon tsaro martani (Ƙarin Table S12).An samo nau'ikan nau'ikan kwayoyin halitta (gungu) ashirin da biyu tare da alaƙa (tabbatacce ko mara kyau) bayanan martaba tsakanin genotypes da bambance-bambancen gwaji. A matsakaita, matakan maganganun kwayoyin halitta suna saukowa cikin tsari 83A: 476> Mandelup> Boregine> Yawan jama'a 22660 (a cikin bambance-bambancen guda biyu, duk da haka, wannan yanayin ya fi ƙarfi a cikin tsire-tsire). A matsakaita, matakan maganganun kwayoyin halitta suna saukowa cikin tsari 83A: 476> Mandelup> Boregine> Yawan jama'a 22660 (a cikin bambance-bambancen guda biyu, duk da haka, wannan yanayin ya fi ƙarfi a cikin tsire-tsire). 83A: 476 > Mandelup > Boregine > Yawan Jama'a 22660 ильнее у контрольных растений). A matsakaita, matakan maganganun kwayoyin sun ragu a cikin tsari 83A: 476> Mandelup> Boregine> Yawan jama'a 22660 (a cikin bambance-bambancen guda biyu, duk da haka, wannan yanayin ya fi ƙarfi a cikin tsire-tsire).平均而言，基因表达水平按83A:476 > Mandelup > Boregine > Yawan jama'a 22660物中更强）.平均 而 言 ， 基因 水平 按 按 83a: 476> mandelup> boregine> yawan jama'a 22660 的 顺序............ 83A: 476 > Mandelup > Boregine > Yawan Jama'a 22660 у контрольных растений). A matsakaita, matakan maganganun kwayoyin sun ragu a cikin jerin 83A: 476> Mandelup> Boregine> Yawan jama'a 22660 (duk da haka, a cikin bambance-bambancen guda biyu, wannan yanayin ya fi ƙarfi a cikin tsire-tsire masu sarrafawa).Alurar riga kafi ya haifar da haɓaka maganganun kwayoyin halitta, musamman a cikin kayayyaki na 18, 19, 14, 6 da 1 (a cikin tsari mai saukowa), ƙa'ida mara kyau (misali modules 9 da 20) ko tare da tasirin tsaka tsaki (misali kayayyaki 11, 22, 8 da 13).Binciken haɓaka kalmar GO (Ƙarin Tebura S13) ya bayyana "GO: 0006952 Amsoshi na Kariya" don tsarin inoculated (18) tare da iyakar kunnawa, gami da nazarin halittu ta qPCR (TanjilG_04706, TanjilG_23384, TanjilG_10657 da TanjilG_23384, TanjilG_10657 da TanjilG_10657 da Tanjil. module (9).Module 18 concentrator (Fig. 8) an gano shi azaman TanjilG_26536 gene wanda ke ɓoye furotin na PR-10-kamar LlR18B, kuma an gano ma'aunin 9 mai maida hankali a matsayin tantanin halitta ta TanjilG_28955 da ke ɓoye furotin II PsbQ.An samo dan takarar anthracnose juriya gene Lanr1, TanjilG_05042, a cikin module 22 (Fig. 9) kuma yana da alaƙa da sharuɗɗan "GO: 0044260 Cellular macromolecular metabolism processes" da "GO: 0006355 Tsarin Rubuce-rubuce, Tsarin DNA" ɗauke da TanjilG_0121 Hub.Halin halittar yana ɓoye ma'aunin matsi na zafi A-4a (HSFA4a).
Binciken cibiyar sadarwa mai nauyi na bayanin haɗin gwiwar kwayoyin halitta tare da madaidaicin sharuddan tsarin nazarin halittu "GO: 0006952 Martanin Tsaro".An sauƙaƙa ligation don haskaka kwayoyin halitta guda huɗu waɗanda qPCR suka bincika (TanjilG_04706, TanjilG_23384, TanjilG_10657 da TanjilG_27015).
Ƙididdigar cibiyar sadarwa mai nauyi na bayanin haɗin gwiwar ƙirar ƙira tare da maƙasudin tsarin tsarin ilimin halitta "GO: 0006355: Tsarin rubutu, gwajin DNA" da ɗaukar ɗan takara juriya na anthracnose Lanr1 TanjilG_05042.An sauƙaƙa ligation don ware jigon TanjilG_05042 da tsakiyar TanjilG_01212 gene.
Binciken juriya na Anthracnose da aka tattara a Ostiraliya ya nuna cewa yawancin cultivars da aka saki da wuri suna da sauƙi;An bayyana Kalya, Coromup da Mandelup a matsayin masu juriya mai matsakaici, yayin da Wonga, Tanjil da 83A:476 aka bayyana su da juriya sosai26,27,31.yana da juriya iri ɗaya, wanda aka sanya Lanr1, kuma Coromup da Mandelup suna da allele daban-daban, waɗanda aka keɓe AnMan10, 26, 39, yayin da Kalya ta wuce ta wani nau'i na daban., Lantarki 2.Binciken juriya na anthracnose a Jamus ya haifar da gano wani layin Bo7212 mai juriya tare da ɗan takara allele banda Lanr1, wanda aka sanya LanrBo36.
Bincikenmu ya nuna ƙarancin mitar (kimanin 6%) na allele na Lanr1 a cikin ƙwayar cuta da aka gwada.Wannan abin lura ya yi daidai da sakamakon gwajin ƙwayar cuta ta Gabashin Turai ta amfani da alamar Anseq3 da Anseq4, wanda ya nuna cewa Lanr1 allele yana cikin layi biyu kawai na Belarushiyanci.Wannan yana nuna cewa har yanzu ba a yi amfani da allele na Lanr1 ta shirye-shiryen kiwo na gida ba, ba kamar a Ostiraliya ba, inda yake ɗaya daga cikin mahimman allele don taimakon kiwo.Wannan na iya zama saboda ƙananan juriya da Lanr1 allele ya bayar a cikin yanayin filin Turai idan aka kwatanta da rahoton Ostiraliya.Bugu da ƙari, nazarin anthracnose a cikin manyan wuraren ruwan sama a Ostiraliya ya nuna cewa amsawar juriya ta hanyar Lanr1 allele bazai yi tasiri ba a yanayin yanayi wanda ke ba da ci gaba da ci gaba da sauri na pathogen19,42.A gaskiya ma, a cikin binciken da aka yi, an kuma lura da wasu alamun anthracnose a cikin genotypes dauke da Lanr1 allele, yana nuna cewa juriya na iya ɓacewa a ƙarƙashin yanayi mafi kyau don ci gaban C. lupini.Bugu da ƙari, fassarori masu kyau na ƙarya na kasancewar alamun Anseq3 da Anseq4, waɗanda suke kusan 1 cm daga wurin Lanr1, suna yiwuwa 28,30,43.
Bincikenmu ya nuna cewa 83A: 476, dauke da Lanr1 allele, ya amsa ga C. lupini inoculation tare da babban sikelin reprogramming reprogramming a farkon binciken lokaci (6 hpi), yayin da a Mandelup, dauke da AnMan allele, transcriptomic martani da aka lura da yawa daga baya.(da 24 a 48 hp).Wadannan bambance-bambance na lokaci-lokaci a cikin martani na tsaro suna da alaƙa da bambance-bambance a cikin alamun cututtuka, suna nuna mahimmancin ganewar asali na farko don samun nasarar amsawa ga juriya.Don cutar da nama na shuka, spores na anthrax dole ne su bi matakai da yawa na haɓakawa akan farfajiyar mai masaukin baki, gami da germination, rarraba tantanin halitta, da samuwar appressorium.Appendage wani tsari ne marar lahani wanda ke mannewa saman mai gida kuma yana sauƙaƙe shiga cikin kyallen da ake watsawa.Don haka, spores na C. gloeosporioides a cikin tsantsar fis ya nuna kashi na farko na tsakiya bayan 75-90 minti na shiryawa, da samuwar kwayar cutar bayan minti 90-120, da kuma dakatarwa bayan 4 hours 45.Mango C. gloeosporioides ya nuna fiye da 40% conidial germination bayan 3 hours na shiryawa da kuma game da 20% samuwar appressors bayan 4 hours.CAP20 na CAP20 da ke da alaka da virulence na C. gloeosporioides ya nuna aikin rubutu a cikin epiphyte-forming conidia bayan 3.5 h incubation a cikin avocado surface wax tare da babban adadin furotin CAP20 bayan 4 h 46 min.Hakazalika, aikin melanin biosynthesis genes a cikin C. trifolii an haifar da shi a lokacin shiryawa na sa'o'i 2 tare da samuwar appressorium bayan sa'a 1.Nazarin leaf kyallen takarda ya nuna cewa strawberries da aka yi wa C. acutatum sun fara kashewa a 8 hpi, yayin da tumatir da aka yi wa C. coccodes sun fara kashewa a 4 hpi48,49.ya yi daidai da ma'aunin lokaci na Colletotrichum spp.tsarin kamuwa da cuta.Amsoshin tsaro da sauri zuwa 83A: 476 suna ba da shawarar shigar da juriya na tsire-tsire da kwayoyin halitta masu haifar da rigakafi (ETI) a cikin wannan layin, yayin da jinkirin martanin Mandelup yana goyan bayan ra'ayin micro-haɗe-haɗe-haɗe-haɗin ƙwayoyin cuta (MTI) hasashe 50. Amsoshi na farko zuwa 83A: 476 da MandelupMatsakaicin juzu'i tsakanin sama-ko ƙasa-kayyade kwayoyin halitta a cikin jinkirin mayar da martani kuma yana goyan bayan wannan ra'ayi, kamar yadda ake ɗaukar ETI azaman haɓakawa da haɓaka amsawar MTI wanda ya ƙare a cikin mutuwar kwayar halitta a wurin kamuwa da cuta, wanda aka sani da girgiza anaphylactic 51,52.
Yawancin kwayoyin halittar da aka danganta ga ma'anar da aka fi sani da Gene Ontology GO: 0006952 "Amsar Tsaro" sune 11 homologues na saƙon azumi mai haifar da damuwa 22 protein (mai kama da SAM22) da kuma manyan abubuwan gina jiki guda bakwai (MLPs).-kamar sunadaran 31, 34, 43 da 423 sun nuna kamanceceniya.SAM22-kamar kwayoyin halitta sun nuna mahimmancin kunnawa wanda ya dade yana dadewa, yana nuna karuwar matakan juriya na anthracnose (83A: 476 da Boregine).Koyaya, kwayoyin halittar MLP-kamar an rage su ne kawai a cikin layin da ke ɗauke da juriyar ɗan takara (83A:476/Lanr1 a 6 hpi da Mandelup/AnMan a 24 hpi).Ya kamata a lura da cewa duk wasu nau'ikan homologues na SAM22 da aka gano sun samo asali ne daga tarin kwayoyin halitta wanda ke da kusan 105 kb, yayin da kwayoyin halittar MLP suka samo asali daga yankuna daban-daban na kwayoyin halitta.An kuma samo haɗin haɗakarwa na irin wannan nau'in SAM22-kamar kwayoyin halitta a cikin bincikenmu na baya na NLL juriya ga maganin Diaporthetoxica, yana nuna cewa suna da hannu a cikin sassan kwance na amsawar tsaro.Hakanan ana samun goyan bayan wannan ƙarshe ta rahotannin ingantaccen amsa na SAM22-kamar kwayoyin halitta don rauni ko jiyya tare da salicylic acid, inducers na fungal, ko hydrogen peroxide.
An nuna kwayoyin halittar MLP masu kama da juna don amsawa ga matsalolin abiotic da biotic iri-iri, gami da ƙwayoyin cuta, ƙwayoyin cuta da cututtukan cututtukan fungal a yawancin nau'ikan shuka55.Hanyoyi na mayar da martani ga wasu mu'amala tsakanin tsire-tsire da ƙwayoyin cuta sun bambanta daga haɓaka mai ƙarfi (watau lokacin da ake kamuwa da auduga tare da Verticillium dahliae) zuwa raguwa sosai (watau bayan kamuwa da itacen apple tare da Alternaria spp.)56,57.An lura da raguwa mai mahimmanci na MLP-kamar 423 gene a lokacin kariyar avocado zuwa F. Niger kamuwa da cuta da kuma lokacin kamuwa da itacen apple Botryosphaeria berengeriana f.cn.piricola da Alternaria alternata su ne apple pathotypes58,59.Bugu da kari, apple calli overexpressing da MLP-kamar 423 gene yana da ƙananan maganganun juriya masu alaƙa da ƙwayoyin cuta kuma sun fi kamuwa da cututtukan fungal59.Bayan Fusarium oxysporum f, MLP-kamar 423 gene kuma an danne a cikin juriya na kowa wake germplasm.cn.Kamuwa da cuta 60.
Sauran membobin dangin PR-10 da aka gano a cikin bincikenmu na RNA-seq sune kwayoyin halittar LlR18A da LlR18B don mayar da martani ga haɓakawa, da kuma haɓakar (1 gene) ko ƙarancin (3 genes) gene don furotin canja wurin lipid DIR1..Bugu da kari, WGCNA tana haskaka halittar LlR18B a matsayin cibiya a cikin wannan tsarin, wanda ke da saurin kamuwa da allurar rigakafi kuma yana dauke da kwayoyin amsawa da yawa.An haifar da kwayoyin LlR18A da LlR18B a cikin ganyen lupine na rawaya don mayar da martani ga kwayoyin cuta na pathogenic, da kuma a cikin NLL mai tushe bayan D. toxica inoculation, yayin da shinkafa homologue na wadannan kwayoyin halitta, RSOsPR10, da sauri ya haifar da kamuwa da cuta na fungal mai yiwuwa ya shiga cikin jasmonic acid siginar hanya, hanyar da ba ta da kwayar cutar D.62. s waɗanda ake buƙata don farkon juriya na tsari (SAR).Tare da haɓaka halayen kariya, ana jigilar furotin DIR1 daga mayar da hankali kan kamuwa da cuta ta cikin phloem don haifar da SAR a cikin gabobin nesa.Abin sha'awa shine, an haifar da kwayar halittar TanjilG_02313 DIR1 sosai a farkon lokaci a cikin layin 84A: 476 da Yawan jama'a 22660, amma juriya na anthracnose ya samu nasara a layin 84A: 476 kawai.Wannan na iya nuna wasu ƙananan aikin kwayar halittar DIR1 a cikin NLL, tun da sauran homologues guda uku sun amsa alluran kawai a cikin 83A: 476 layin a 6 hpi, kuma wannan amsa ya koma ƙasa.
A cikin bincikenmu, mafi yawan abubuwan da suka fi dacewa da tsarin ilimin halitta da ake kira "GO: 0055114 Redox tsari" sune furotin P450 cytochrome, peroxidase, linoleic acid 9S- / 13S-lipoxygenase, da 1-aminocyclopropane-1-carboxylic acid oxidase.Bugu da kari, WGCNA ta mu ta ayyana homologue na HSFA4a a matsayin cibiya mai dauke da kayayyaki irin su dan takarar juriya na Lanr1 TanjilG_05042.HSFA4a wani ɓangare ne na ƙa'idodin dogaro da redox na rubutun nukiliya a cikin tsirrai.
Cytochrome P450 sunadaran ne oxidoreductases cewa catalyze NADPH da / ko O2-dogara hydroxylation halayen a primary da sakandare metabolism, ciki har da metabolism na xenobiotics, kazalika da hormones, m acid, sterols, cell bango aka gyara, biopolymers, da biosynthesis na m mahadi 69. A cikin aikin chromto00 ya rage daga chromocyte 0. 6 log2 (canji ninki) zuwa 5.7 saboda ɗimbin sauye-sauye na homologues (37) da bambance-bambance a cikin tsarin amsawa tsakanin takamaiman kwayoyin halitta, yana nuna bita na sama..Yin amfani da bayanan RNA-seq kawai don haɓaka aikin nazarin halittu na kwayoyin halittar NLL a cikin irin wannan babban rukunin furotin zai zama hasashe sosai.Duk da haka, yana da mahimmanci a lura cewa wasu kwayoyin cytochrome P450 suna da alaƙa da haɓakar juriya ga fungi ko kwayoyin cuta, ciki har da gudummawa ga rashin lafiyar jiki69,70,71.
Class III peroxidases ne multifunctional shuka enzymes da hannu a cikin wani m kewayon na rayuwa tafiyar matakai a lokacin shuka girma da kuma ci gaban shuka, kazalika da mayar da martani ga muhalli danniya kamar salinity, fari, high haske tsanani, da pathogen attack72.Peroxidases suna shiga cikin hulɗar nau'in tsire-tsire masu yawa tare da Anthracis, ciki har da Stylosanthes humilis da C. gloeosporioides, Lens culinaris da C. truncatum, Phaseolus vulgaris da C. lindemuthianum, Cucumis sativus da C. lagenarium73,764.75.Amsar tana da sauri sosai, wani lokacin ma a 4 HPI, kafin naman gwari ya shiga cikin shuka73.Halin peroxidase kuma ya amsa D. toxica NLL inoculation.Baya ga ayyukansu na yau da kullun don daidaita fashewar oxidative ko kawar da damuwa na oxidative, peroxidases na iya tsoma baki tare da haɓakar ƙwayoyin cuta ta hanyar ƙirƙirar shingen jiki dangane da ƙarfafa bangon tantanin halitta yayin lignification, subunit ko haɗin giciye na takamaiman mahadi.Ana iya danganta wannan aikin a cikin silico zuwa ga TanjilG_03329 gene wanda ke ɓoye wani lignin-forming anion peroxidase wanda aka haɓaka sosai a cikin bincikenmu a cikin 83A: 476 layin juriya a 6 HPI, amma ba a cikin wasu nau'ikan da wuraren lokaci waɗanda basu amsa ba.
9S-/13S-lipoxygenase na linoleic acid shine mataki na farko a cikin hanyar oxidative na lipid biosynthesis78.Samfuran wannan hanyar suna da ayyuka da yawa a cikin kariyar shuka, gami da ƙarfafa bangon tantanin halitta ta hanyar samar da adibas na callose da pectin, da ka'idojin damuwa na oxidative ta hanyar samar da nau'in oxygen mai amsawa79,80,81,82,83.A cikin binciken yanzu, an canza bayanin linoleic acid 9S-/13S-lipoxygenase a cikin kowane nau'i, amma a cikin yawan jama'a 22660, haɓakawa ya rinjayi a lokuta daban-daban, yayin da a cikin nau'o'in da ke dauke da Lanr1 mai tsayayya da AnMan allele, yana jaddada bambancin nau'i na oxylipins a tsakanin wadannan gefuna na gefuna.
Homologue na 1-aminocyclopropane-1-carboxylate oxidase (ACO) an inganta shi sosai (genes 9) ko raguwa (2 genes) lokacin da aka shafe shi da lupine.Tare da keɓancewa guda biyu, duk waɗannan martanin sun faru a 6 hp.da 83:476.Halin enzymatic wanda furotin ACO ke shiga tsakani shine matakin iyakance adadin ethylene don haka yana da tsari sosai84.Ethylene hormone ne na tsire-tsire wanda ke taka rawa iri-iri a cikin daidaita tsarin ci gaban shuka da kuma mayar da martani ga yanayin damuwa da ƙwayoyin cuta.Shigar da rubutun ACO da kunna hanyar siginar ethylene suna da hannu wajen haɓaka juriyar shinkafa ga naman gwari na hemibiotrophic oryzae oryzae ta hanyar daidaita samar da nau'in oxygen mai amsawa da phytoalexins.Tsarin kamuwa da cuta mai kama da ganye da aka samu tsakanin M. oryzae da C. lupini88,89, a kan bangon babban haɓakar ACO homologues a cikin layin 83A: 476 da aka ruwaito a cikin wannan binciken, yana canza yiwuwar ba da juriya ga NLL anthracnose Ethylene alama ce ta tsakiya a cikin hanyoyin ƙwayoyin cuta.
A cikin binciken da ake yi yanzu, an lura da babban kashe kwayoyin halittar da ke da alaƙa da photosynthesis a 6 hpi a cikin 83A: 476 da kuma a 48 hpi a cikin Mandeloop da yawan 22660.Girma da ci gaban waɗannan canje-canje sun yi daidai da matakin.An lura da juriya na Anthracnose a cikin wannan gwaji.Kwanan nan, an ba da rahoto mai ƙarfi da farkon danniya na rubuce-rubuce masu alaƙa da photosynthesis a cikin nau'ikan nau'ikan hulɗar ƙwayoyin cuta da yawa, gami da ƙwayoyin cuta da fungi.Gaggauta (daga 2 HPI a wasu hulɗar) da ƙaddamar da ƙwayoyin halittar da ke da alaƙa da photosynthesis a cikin martani ga kamuwa da cuta na iya haifar da rigakafi na shuka dangane da jigilar nau'in oxygen mai amsawa da hulɗar su tare da hanyar salicylic acid don daidaita yanayin rashin lafiyan 90,94.
A ƙarshe, hanyoyin mayar da martani na tsaro da aka ba da shawarar don mafi girman layin juriya (83A: 476) sun haɗa da saurin ganewar ƙwayoyin cuta ta hanyar R gene (wataƙila TIR-NBS-LRR TanjilG_05042) da rashin lafiyar salicylic acid da siginar ethylene, wanda ya biyo bayan kafa SAR mai tsayi.aikin yana goyan bayan furotin DIR-1.Ya kamata a lura cewa lokacin biotrophic don kamuwa da cutar C. lupini yana da ɗan gajeren lokaci (kimanin kwanaki 2), sannan kuma ci gaban necrotic95.Canje-canjen tsakanin waɗannan matakai na iya haɗawa da necrosis da kuma bayyanar da sunadaran ethylene-inducible wanda ke aiki a matsayin abubuwan da ke haifar da halayen haɓakawa a cikin tsire-tsire masu tsire-tsire.Saboda haka, taga lokacin don nasarar kama C. lupini a matakin biotrophic yana da kunkuntar sosai.Sake fasalin kwayoyin halittar da ke hade da redox da photosynthesis da aka lura a cikin 83A: 476 a 6 hpi ya dace da ci gaban hyphae na fungal kuma yana ba da sanarwar ci gaba da nasarar amsawar kariya a matakin biotrophic.Amsoshin fassarar Mandelup da yawan mutanen 22660 na iya yin jinkiri sosai don kama naman gwari kafin su canza zuwa ci gaban necrotic, duk da haka, Mandelup na iya zama mafi inganci fiye da yawan 22660 saboda ƙayyadaddun ƙayyadaddun tsari na furotin PR-10 yana haɓaka juriya a kwance.
ETI, wanda ke gudana ta hanyar canonical R gene, ya bayyana azaman hanyar gama gari don juriyar wake ga anthracnose.Don haka, a cikin samfurin legume Medicago truncatula, juriya ga anthracnose yana ba da shi ta hanyar RCT1 gene, memba na TIR-NBS-LRR97 shuka R gene class.Wannan kwayar halitta kuma tana ba da juriya mai faɗin anthracnose a cikin alfalfa lokacin da aka canza shi zuwa tsire-tsire masu saurin kamuwa.A cikin wake na kowa (P. vulgaris), an gano fiye da dozin biyu na juriya na anthracnose zuwa yau.Wasu daga cikin waɗannan kwayoyin halitta ana samun su a yankuna da ba su da kowane nau'in halittar R, duk da haka wasu da yawa suna nan a gefen chromosomes masu ɗauke da tarin kwayoyin halittar NBS-LRR, gami da TIR-NBS-LRRs99.Binciken SSR na genome-fadi ya kuma tabbatar da haɗin gwiwar kwayar halittar NBS-LRR tare da juriya na anthracnose a cikin wake na kowa.Hakanan an samo asalin halittar R na canonical a cikin yankin genomic yana ɗauke da babban wurin juriya na anthracnose a cikin farin lupine 101.
Ayyukanmu sun nuna cewa amsawar juriya nan da nan, wanda aka kunna a farkon matakin kamuwa da cuta (zai fi dacewa ba daga baya ba fiye da 12 hpi), da kyau yana kare lupine mai kunkuntar daga anthracnose wanda ya haifar da naman gwari na Collelotrichum lupini.Yin amfani da jerin abubuwan da aka samu mai girma, mun nuna bambancin bayanin bayanan martaba na kwayoyin juriya na anthracnose a cikin tsire-tsire na NLL wanda Lanr1 da AnMan juriya suka yi.Nasarar tsaro ya ƙunshi ƙirƙira a hankali tsara kwayoyin halitta don sunadaran da ke da hannu a redox, photosynthesis, da pathogenesis cikin sa'o'i na farkon tuntuɓar shuka da cuta.Irin wannan halayen kariya, amma jinkirta cikin lokaci, ba su da tasiri sosai wajen kare tsirrai daga cututtuka.Juriya na Anthrax wanda ke yin sulhu ta hanyar Lanr1 gene yayi kama da irin saurin amsawar kwayar halittar R (maganin rigakafi mai haifar da tasiri), yayin da kwayar halittar AnMan ta fi dacewa ta ba da amsa a kwance (kayan rigakafi da ke haifar da tsarin kwayoyin halitta mai alaka da microbe), yana samar da matsakaicin matakin dorewa.
Layukan NLL 215 da aka yi amfani da su don tantance alamun anthracnose sun ƙunshi nau'ikan cultivars 74, layin 60 da aka samu ta hanyar hayewa ko kiwo, 5 mutants, da 76 na daji ko na asali germplasms.Layukan sun fito ne daga kasashe 17, galibi daga Poland (58), Spain (47), Jamus (27), Australia (26), Rasha (19), Belarus (7), Italiya (5) da sauran layin.daga kasashe 10.Saitin kuma ya haɗa da layukan juriya: 83A: 476, Tanjil, Wonga ɗauke da Lanr1 allele, da Mandelup ɗauke da allelen AnMan.An samo layukan daga Rukunin Bayanan Halitta na Lupine na Turai wanda Poznań Plant Breeding Ltd., Wiatrowo, Poland ke kula da shi (Ƙarin Teburin S1).
An girma tsire-tsire a ƙarƙashin yanayin sarrafawa (photoperiod 16 hours, zazzabi 25 ° C a rana da 18 ° C da dare).An yi nazarin kwafin halittu guda biyu.An ware DNA daga ganyen makonni uku ta amfani da DNeasy Plant Mini Kit (Qiagen, Hilden, Jamus) bisa ga yarjejeniya.An kimanta inganci da tattarawar DNA ta keɓe ta hanyoyin spectrophotometric (NanoDrop 2000; Thermo Fisher Scientific, Waltham, MA, Amurka).An yi la'akari da alamar AnManM1 da ke nuna alamar juriya na anthracnose AnMan (wanda aka samo daga cv. Mandelup) da alamomin Anseq3 da Anseq4 flanking gene Lanr1 (wanda aka samo daga cv. Tanjil) 11,26,28.Homozygotes na allele masu juriya an saka su a matsayin “1″, mai saukin kamuwa – kamar “0″, da heterozygotes – kamar 0.5.
Dangane da sakamakon bincike don alamomin AnManM1, AnSeq3 da AnSeq4 da wadatar tsaba don gwaje-gwaje na ƙarshe na ƙarshe, an zaɓi layin 50 NLL don phenotyping juriya na anthracnose.An gudanar da bincike a cikin kwafi a cikin injin da aka sarrafa na kwamfuta tare da lokacin daukar hoto na sa'o'i 14 tare da zafin jiki na 22 ° C a rana da 19 ° C da dare.Ana goge iri (yanke rigar iri a gefen tayin tare da kaifi mai kaifi) kafin shuka don hana zurfafa zurfafawa saboda gashin iri yana da ƙarfi da kuma tabbatar da fitowar iri ɗaya.An shuka tsire-tsire a cikin tukwane (11 × 11 × 21 cm) tare da ƙasa mara kyau (TS-1 REC 085 Medium Basic, Klasmann-Deilmann Polska, Warsaw, Poland).An gudanar da inoculation tare da nau'in Colletotrichum lupini Col-08, wanda aka girma a cikin 1999 daga mai tushe na tsire-tsire masu tsire-tsire masu tsire-tsire da aka girma a cikin wani fili a Verzhenitsa, Greater Poland (52° 27′ 42″ N 17° 04′ 05″ E).Samu yanki.An haɓaka keɓancewar a cikin matsakaicin SNA a 20 ° C. ƙarƙashin hasken baƙar fata na kwanaki 21 don haifar da sporulation.Makonni hudu bayan shuka, lokacin da tsire-tsire suka kai matakin ganye na 4-6, an gudanar da allurar ta hanyar fesa tare da dakatarwar conidia a matakin 0.5 x 106 conidia a kowace ml.Bayan yin allurar, an ajiye tsire-tsire a cikin duhu na tsawon sa'o'i 24 a yanayin zafi na kusan 98% da zafin jiki na 25 ° C don sauƙaƙe germination na conidia da tsarin kamuwa da cuta.An shuka tsire-tsire a ƙarƙashin yanayin hoto na awa 14 a 22 ° C rana / 19 ° C da dare da 70% zafi.An yi macijin cutar kwanaki 22 bayan allurar rigakafi kuma ya kasance daga 0 (na rigakafi) zuwa 9 (mai saurin kamuwa da cuta) dangane da kasancewar ko rashin raunin necrotic akan mai tushe da ganye.Bugu da ƙari, bayan zura kwallo, an auna nauyin tsire-tsire.Dangantaka tsakanin alamomin genotypes da nau'ikan cututtukan cututtuka an ƙididdige su azaman ma'ana guda biyu masu alaƙa (rashin alamun heterozygous a cikin saitin layi don nazarin phenotype juriya na anthracnose).