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Angustifolius lupine (NLL, Lupinus angustifolius L.) ke semela sa linaoa se sebelisetsoang tlhahiso ea lijo le ntlafatso ea mobu.Katoloso ea lefats'e ea NLL joalo ka sejalo e hohetse li-fungus tse ngata tsa pathogenic, ho kenyelletsa le lupine anthracnose, e bakang lefu le senyang la anthracnose.Li-alleles tse peli, Lanr1 le AnMan, tse fanang ka khanyetso e eketsehileng, li sebelisitsoe ho ikatisa ha NLL, empa mekhoa ea limolek'hule e ntse e sa tsejoe.Phuputsong ena, ho ile ha sebelisoa matšoao a Lanr1 le AnMan ho hlahloba lisampole tsa NLL tsa Europe.Teko ea ente tikolohong e laoloang e netefalitse ho sebetsa ha bafani ka bobeli ba hanyetsang.Phapang ea polelo ea liphatsa tsa lefutso e ne e etsoa methating e hanyetsanang le baemeli le e hlaselehang habonolo.Ho hanyetsa anthracnose ho ne ho amahanngoa le tlhaloso e feteletseng ea mantsoe a liphatsa tsa lefutso "GO: 0006952 Defense Response", "GO:0055114 Redox Process", le "GO:0015979 Photosynthesis".Ho phaella moo, mola oa Lanr1 (83A: 476) o bontšitse phetoho e kholo ea transcriptome ka potlako ka mor'a ho kenngoa, ha mela e meng e bontša ho lieha ho arabela ka lihora tse ka bang 42.Likarabo tsa tšireletso li amahanngoa le liphatsa tsa lefutso tsa TIR-NBS, CC-NBS-LRR le NBS-LRR, liprotheine tsa 10 tse amehang ho pathogenesis, liprotheine tsa ho fetisa lipid, endoglucan-1,3-β-glucosidase, liprotheine tsa marako a glycine tse ngata, le liphatsa tsa lefutso tse tsoang tseleng e sebetsang ea oksijene.Likarabo tsa pele ho 83A: 476, ho kenyelletsa le khatello e hlokolosi ea liphatsa tsa lefutso tse amanang le photosynthesis, li tsamaellana le ts'ireletso e atlehileng nakong ea kholo ea limela ea fungal biology, e fana ka maikutlo a hore phello e baka ts'ireletso ea 'mele.Boitšoaro ba Mandeloop bo fokotseha, joalo ka ho hula ka kakaretso.
Lupine e mahlaku a moqotetsane (NLL, Lupinus angustifolius L.) ke lijo-thollo tse nang le protheine e ngata e tsoang sebakeng se ka bophirimela sa Mediterranean1,2.Hajoale e lengoa joalo ka sejalo sa lijo tsa liphoofolo le batho.E boetse e nkoa e le moiteli o motala mekhoeng ea ho potoloha ha lijalo ka lebaka la ho lokisa naetrojene ka symbiotic nitrogen fixing bacteria le ntlafatso ea kakaretso ea mobu oa mobu.NLL e bile le ts'ebetso e potlakileng ea ho rua malapeng lekholong le fetileng la lilemo mme e ntse e le tlas'a khatello e phahameng ea ho ikatisa3,4,5,6,7,8,8,9,10,11,12.Ka temo e atileng ea NLL, tatellano ea li-fungus tsa pathogenic e ile ea hlahisa libaka tse ncha tsa temo mme tsa baka mafu a macha a senyang lijalo. Ntho e tsotehang ka ho fetisisa ho lihoai tsa lupine le bahlahisi e ne e le ponahalo ea anthracnose, e bakoang ke fungus ea pathogenic, Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Ntho e tsotehang ka ho fetisisa ho lihoai tsa lupine le bahlahisi e ne e le ponahalo ea anthracnose, e bakoang ke fungus ea pathogenic, Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Наиболее примечательным для фермеров и селекционеров люпина было появление антракноза, вызванного патогенным грибком Colletotrichum, Fellerdor Se ileng sa hlokomeleha haholo ho lihoai le balemi ba lupine ke ho hlaha ha anthracnose e bakoang ke fungus ea pathogenic Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13.对于羽扇豆农民和饲养者來说，最引人注目的是炭疽病的起现，它是由病原真菌Colletotrichum lupini对于羽扇豆农民和饲养者來说，最引人注目的是炭疽病的出现，它是由病原真菌Colletotrichum lupini). Наиболее поразительным для фермеров и селекционеров люпина является появление антракноза, вызываемого патогененым гриtricбком Colleum1 . Ntho e khahlang ka ho fetesisa ho lihoai le balemi ba lupine ke ho hlaha ha anthracnose e bakoang ke fungus ea pathogenic Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13.Litlaleho tsa pele tsa lefu lena li tsoa Brazil le United States, tse nang le matšoao a tloaelehileng a hlahang ka 1912 le 1929 ka ho latellana.Leha ho le joalo, ka mor’a lilemo tse ka bang 30, kokoana-hloko ena e ile ea khethoa e le Colletotrichum gloeosporioides (Penz.) Penz. & Sacc., teleomorph Glomerella cingulata (Stoneman) Spauld. & Sacc., teleomorph Glomerella cingulata (Stoneman) Spauld. & Sacc., телеоморф Glomerella cingulata (Stoneman) Spauld. & Sacc., teleomorph of 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 Targeted Morphology. & H. Schrenk,. & H. Schrenk, .le H. Schrenk. & H.施伦克，. & H.施伦克，.le H. Schlenk, .Phenotyping ea pele ea lefu e entsoeng bohareng ba lekholo la bo20 la lilemo e bontšitse khanyetso e itseng ho NLL le li-accessions tsa yellow lupine (L. luteus L.), empa li-accessions tsohle tse tšoeu tsa lupine (L. albus L.) tse lekiloeng li ne li le bonolo haholo15,16.Liphuputso li bontšitse hore nts'etsopele ea anthracnose e amahanngoa le ho eketseha ha pula (mongobo oa moea) le mocheso (ka bophara ba 12-28 ° C), e leng se lebisang ho tlōlo ea ho hanyetsa mocheso o phahameng17, 18. Ha e le hantle, nako e hlokahalang bakeng sa hore conidia e mele le lefu lena le qala , e ne e le khutšoanyane ka makhetlo a mane ho 24 ° C (lihora tse 12) ho feta maemo a phahameng a 12 ° C (lihora tse 12).Kahoo, ho futhumala ha lefatše ho ntseng ho tsoela pele ho lebisitse ho ata ha anthracnose.Leha ho le joalo, lefu lena le ile la hlokomeloa Fora (1982) le Ukraine (1983) e le sesupo sa tšokelo e neng e le haufi, empa ho bonahala eka indasteri ea lupine e ile ea hlokomolohuoa ka nako eo20,21.Lilemo tse ’maloa hamorao, lefu lena le senyang le ile la ata lefatšeng lohle ’me la ama linaha tse khōlō tse hlahisang lupine tse kang Australia, Poland le Jeremane22,23,24.Ka mor'a ho qhoma ha anthracnose bohareng ba lilemo tsa bo-1990, tlhahlobo e pharaletseng e ile ea fella ka ho tsebahatsoa ha bafani ba 'maloa ba hanyetsang lisampole tsa NLL19.Ho hanyetsa ha NLL ho anthracnose ho laoloa ke li-allele tse peli tse arohaneng tse fumanehang mehloling e fapaneng ea likokoana-hloko: Lanr1 ho mofuta oa Tanjil le Wonga le AnMan mokhahlelong.Mandalay 25, 26. Li-allele tsena li tlatsana le matšoao a molek'hule a tšehetsang khetho ea likokoana-hloko tse hanyetsanang le mananeo a ho ikatisa25,26,27,28,29,30.Mohala o hanyetsanang oa ho ikatisa oa 83A: 476 o jereng Lanr1 allele o ile oa tšeloa ka mohala oa hlaha oa P27255 ho fumana palo ea batho ba RIL e arohaneng bakeng sa ho hanyetsa anthracnose, e leng se entseng hore ho khonehe ho abela sebaka sa Lanr1 ho chromosome NLL-1131, 32, 33. moralo, NLL e senotse sebaka sa alleles tse tharo ho chromosome e le 'ngoe (NLL-11), empa maemong a fapaneng29,34,35.Leha ho le joalo, ka lebaka la palo e fokolang ea li-RIL le sebaka se seholo sa liphatsa tsa lefutso pakeng tsa matšoao le li-alleles tse lumellanang, ha ho liqeto tse ka tšeptjoang tse ka etsoang mabapi le liphatsa tsa tsona tsa lefutso.Ka lehlakoreng le leng, ts'ebeliso ea liphatsa tsa lefutso ho lupin e thata ka lebaka la matla a bona a tlase haholo a ho nchafatsa, e leng se etsang hore liphatsa tsa lefutso li be boima37.
Tsoelo-pele ea likokoana-hloko tse ruuoang lapeng tse nang le allele e lakatsehang sebakeng sa homozygous, joalo ka 83A: 476 (Lanr1) le Mandelup (AnMan), e butse monyetla oa ho ithuta ho hanyetsa anthracnose ka lebaka la ho ba teng ha metsoako e hanyetsanang ea alleles ho baahi ba hlaha.Menyetla ea mekhoa ea limolek'hule.Bapisa likarabo tsa ts'ireletso tse hlahisoang ke li-genotype tse itseng.Phuputso ena e ile ea hlahloba karabelo ea pele ea transcriptome ea NLL ho ente ea C. lupini.Taba ea pele, karolo ea European NLL germplasm e nang le mela e 215 e ile ea hlahlojoa ho sebelisoa matšoao a molek'hule a tšoaeang Lanr1 le AnMan alleles.Anthracnose phenotyping e ile ea etsoa ka mela ea 50 NLL, e neng e khethiloe pele bakeng sa matšoao a limolek'hule, tlas'a maemo a laoloang.Ho ipapisitsoe le liteko tsena, ho ile ha khethoa mela e mene e fapaneng ka khanyetso ea anthracnose le sebopeho sa allelic sa Lanr1/AnMan bakeng sa profiling e fapaneng ea liphatsa tsa lefutso ho sebelisoa mekhoa e 'meli e tlatselletsang: tatellano e phahameng ea RNA le quantification ea nako ea nnete ea PCR.
Ho hlahlojoa ha sete ea NLL germplasm (N = 215) e nang le matšoao a Lanr1 (Anseq3 le Anseq4) le AnMan (Anseq4) le AnMan (AnManM1) e bonts'itse hore ke mola o le mong feela (95726, haufi le Salamanca-b) o matlafatsang "ho hanyetsa" allele bakeng sa matšoao ohle, athe "Presceptible" tsohle li ka fumanoa. (~ 73.5%) mela.Methalo e leshome le metso e meraro e hlahisitse alleles tse peli tse "hanellang" tsa lesupa la Lanr1, 'me mela e 8 e hlahisitse alleles "tse hanyetsanang" tsa Lanr1.lesupa."Khanyetso" allele ea lesupa la AnMan (Tafole ea Tlatsetso S1).Mela e 'meli e ne e le heterozygous bakeng sa lesupa la Anseq3 le heterozygous e le' ngoe bakeng sa lesupa la AnManM1.Mehala ea 42 (19.5%) e ne e e-na le mekhahlelo e fapaneng ea Anseq3 le Anseq4 alleles, e bontšang hore ho na le maqhubu a phahameng a ho kopanya pakeng tsa li-loci tsena tse peli.Anthracnose phenotypes tlas'a maemo a laoloang (Supplementary Table S2) e senotse ho fapana ha ho hanyetsa ha li-genotype tse lekiloeng, tse neng li bonahala ka ho teba ha anthracnose.Phapang ea lintlha tse tloaelehileng e ne e tloha ho 1.8 (e hanyetsanang ka mokhoa o itekanetseng) ho ea ho 6.9 (e hlaselehang habonolo) le phapang ea boima ba semela e tloha ho 0.62 (e hlaselehang habonolo) ho ea ho 4.45 g (e hanyetsanang). Ho ne ho e-na le kamano e kholo pakeng tsa litekanyetso tse hlokometsoeng liphetolelong tse peli tsa teko (0.51 bakeng sa lintlha tse matla tsa lefu, P = 0.00017 le 0.61 bakeng sa boima ba limela, P <0.0001) hammoho le pakeng tsa mekhahlelo ena e 'meli (- 0.59 le - 0.77, P0100). Ho ne ho e-na le kamano e kholo pakeng tsa litekanyetso tse hlokometsoeng liphetolelong tse peli tsa teko (0.51 bakeng sa lintlha tse matla tsa lefu, P = 0.00017 le 0.61 bakeng sa boima ba semela, P <0.0001) hammoho le pakeng tsa liparamente tsena tse peli (- 0.59 le - 0.0000 <1). Выявлена ​​достоверная корреляция между значениями, наблюдаемыми в двух повторностях эксперимента (0,51 для баллов тяжезд00, 1 = 1 = массы растения, P < 0,0001), а также между этими двумя параметрами (-0,59 и -0,77, Р < 0,0001) 0,0001). Ho ile ha fumanoa kamano ea bohlokoa pakeng tsa litekanyetso tse hlokometsoeng ka makhetlo a mabeli a teko (0.51 bakeng sa lintlha tse matla tsa lefu, P = 0.00017 le 0.61 bakeng sa boima ba semela, P <0.0001), hammoho le pakeng tsa mekhahlelo ena e 'meli (- 0.59 le -0.77, P010) 0.0).在两次重复实验中观察到的值之间存在显着相关性（疾病严重程度评分為0.51，P = 110.000 = 0.00. 0.0001）以及這两个参数之间（- 0.59 和- 0.77，P <0.0001).在在 两两 的 的 的 的 的 的 的 存在 存在 相关性 相关性 相关性 分为 相关性 分为 分为 分为 分为 分为 分为 分为 分为 分为 分为 个 个 个 个 个 个 个 之间 个 之间 Наблюдалась значительная корреляция между значениями, наблюдаемыми в двух повторностях (оценка тяжести заболевания 0,51, 10 = 10 = P <0,0001), и между этими двумя параметрами (-0,59 и -0,0001) 0,77, P <0,0001. Ho ne ho e-na le kamano e kholo pakeng tsa litekanyetso tse hlokometsoeng ka makhetlo a mabeli (lintlha tse matla tsa lefu 0.51, P = 0.00017 le boima ba semela 0.61, P <0.0001) le pakeng tsa liparamente tsena tse peli (-0.59 le -0 .0001) 0.77, P <0.000. ).Matšoao a tloaelehileng a bonoang limeleng tse nang le ts'oaetso a kenyelletsa kinking le ho sotha ha kutu ho tšoana le sebopeho sa "seqha sa molisa", se lateloa ke maqeba a oval a nang le li-orange / pink sporozoites (Fig. 1 ea Tlatsetso).Likhetho tsa Australia tse jereng liphatsa tsa lefutso tsa Lanr1 (83A:476 le Tanjil) le AnMan (Mandelup) li hanyetsana ka mokhoa o itekanetseng, 0.0331 le 0.0036).Methalo e meng e nang le "resistant" Lanr1 le/kapa AnMan alleles e bontša matšoao a lefu lena.
Hoa thahasellisa hore mela e 'maloa ea NLL e haelloang ke "resistant" marker allele e senotse boemo bo phahameng ba ho hanyetsa anthracnose (e ka bapisoang kapa e phahameng ho feta ea Lanr1 kapa AnMan genotypes), e kang Boregine (P value <0.0001 bakeng sa liparamente ka bobeli), Bojar (P boleng <0.0001000 boleng ba B-P9 bakeng sa palo ea B / P9. 0.0001 bakeng sa lintlha le tse seng bohlokoa bakeng sa boima ba 'mele). Hoa thahasellisa hore mela e 'maloa ea NLL e haelloang ke "resistant" marker allele e senotse boemo bo phahameng ba ho hanyetsa anthracnose (e ka bapisoang kapa e phahameng ho feta ea Lanr1 kapa AnMan genotypes), e kang Boregine (P value <0.0001 bakeng sa liparamente ka bobeli), Bojar (P boleng <0.0001000 boleng ba B-P9 bakeng sa palo ea B / P9. 0.0001 bakeng sa lintlha le tse seng bohlokoa bakeng sa boima ba 'mele). Интересно, что несколько линий NLL, лишенных какого-либо «резистентного» маркерного аллеля, показали высокий уровень е высокий, чем для генотипов Lanr1 или AnMan), таких как Boregine (значение P <0,0001 для обоих параметров), Bojar (значение P < 0,0001 0,0001 для обоих параметров), Bojar стения) и популяции B-549/79b (значение P <0,0001 для оценки и незначимо для массы). Hoa thahasellisa hore mela e mengata ea NLL e haelloang ke "resistant" marker allele e bontšitse boemo bo phahameng ba ho hanyetsa anthracnose (e bapisoang kapa e phahameng ho feta ea Lanr1 kapa AnMan genotypes), e kang Boregine (P boleng <0.0001 bakeng sa liparamente ka bobeli), Bojar (P boleng <0.0001 bakeng sa palo ea batho ba 7 / 9 bakeng sa 0.0001 bakeng sa 0.0001 le B900 bakeng sa semela sa 9b-9). P boleng <0.0001 bakeng sa tlhahlobo mme eseng ea bohlokoa bakeng sa boima ba 'mele).有趣的是，一些缺乏任何“抗性”标记等位基因的NLL 系显示出高水平的炭疽病抗性（帎LanAn1更高），例如Boregine（两个参数的P 值<0.0001）、Bojar（P 值<得分為0.0001，植物重和量為0.0001）B71566001（90001值< 0.0001, 重量不显着）. Hoa thahasellisa hore litsamaiso tse ling tsa NLL tse se nang matšoao a "antigenic" li bonts'a khanyetso e phahameng e holimo (e lekanang le Lanr1 kapa AnMan liphatsa tsa lefutso kapa tse phahameng), tse kang Boregine (ka bobeli li-parameter P <0.0001), Bojar (P boleng <0.0001, boima ba semela 0.001) le boima ba 4 /0 0 000 0.5 bohlokoa <0 0 0 0 000 . Интересно, что некоторые линии NLL, лишенные каких-либо маркерных аллелей «резистентности», показали высокие уровни устойчинизсти чем у генотипов Lanr1 или AnMan), такие как Boregine (значение P для обоих параметров <0,0001), Bojar (значение P <0,0001, масса раст00лц5) и 9 енка P-значение <0,0001, масса незначительна). Hoa thahasellisa hore mela e meng ea NLL e haelloang ke "resistance" marker alleles e bontšitse litekanyetso tse phahameng tsa khanyetso ea anthracnose (e bapisoang kapa e phahameng ho feta Lanr1 kapa AnMan genotypes), joalo ka Boregine (P bohlokoa bakeng sa parameters ka bobeli <0.0001), Bojar (P-boleng <0.0000.701, P-701, P-value <0.00001, P-901, P-701, P-value <0.0001,901,901, P-value) <0.0001, boima ha bo bohlokoa).Ketsahalo ena e fana ka maikutlo a monyetla oa hore ho be le mohloli oa liphatsa tsa lefutso oa khanyetso, o hlalosang khaello e hlokomelehang ea kamano pakeng tsa li-marker genotypes le mafu a phenotypes (P boleng ho tloha ~ 0.42 ho ~ 0.98).Kahoo, tlhahlobo ea Kolmogorov-Smirnov e bonts'itse hore lintlha tsa ho hanyetsa anthracnose li ne li batla li ajoa bakeng sa lintlha (P-values ​​0.25 le 0.11) le boima ba limela (P-values ​​0.47 le 0.55), ho fana ka maikutlo a hore ke nahana hore ho na le li-alleles tse ngata ho feta Lanr1 le AnMan.
Ho itšetlehile ka liphello tsa tlhahlobo ea ho hanyetsa anthracnose, mela ea 4 e khethiloe bakeng sa tlhahlobo ea transcriptome: 83A: 476, Boregine, Mandelup, le Population 22660. Mela ena e ile ea hlahlojoa hape bakeng sa ho hanyetsa koatsi litekong tsa inoculation ka tatellano ea RNA, hafeela e ne e tšoana le tekong e fetileng.Maemo a lintlha e ne e le a latelang: Boregin (1.71 ± 1.39), 83A: 476 (2.09 ± 1.38), Mandelup (3.82 ± 1.42) le baahi ba 22660 (6.11 ± 1.29 ).
Protocol ea Illumina NovaSeq 6000 e fihletse karolelano ea lipara tsa Mread tse 40.5 ka mohlala (29.7 ho isa ho 54.4 Mreads) (Tlaleho ea Tlatsetso ea S3).Lintlha tsa ho tsamaisana ka tatellano ea litšupiso li tloha ho 75.5% ho isa ho 88.6%.Kakaretso ea khokahano ea lintlha tse baloang lipakeng tsa mefuta ea liteko lipakeng tsa likopi tsa baeloji ho tloha ho 0.812 ho isa ho 0.997 (e bolela 0.959). Ho tsoa ho liphatsa tsa lefutso tsa 35,170 tse hlahlobiloeng, 2917 ha ea ka ea senola, 'me liphatsa tse ling tsa 4785 li ile tsa hlahisoa ka mokhoa o sa tsotelleng (motheo o bolela <5). Ho tsoa ho liphatsa tsa lefutso tsa 35,170 tse hlahlobiloeng, 2917 ha ea ka ea senola, 'me liphatsa tse ling tsa 4785 li ile tsa hlahisoa ka mokhoa o sa tsotelleng (motheo o bolela <5). Из 35 170 проанализированных генов 2917 не проявляли экспрессии, а остальные 4785 генов экспрессировались на ненезнабрессии. Har'a liphatsa tsa lefutso tsa 35,170 tse hlahlobiloeng, 2917 ha ea ka ea bontša polelo, 'me liphatsa tsa lefutso tse setseng tsa 4785 li ile tsa hlahisoa ka mokhoa o sa tsotelleng (motheo o bolela <5).在分析的35,170 个基因中，2917 个沒有表达，其他4785 个基因的表达可以忽略不计（基本平均倉。35,170 Из 35 170 проанализированных генов 2917 не экспрессировались, а остальные 4785 генов имели незначительную экспрессировались, а остальные 4785 генов имели незначительную экспрессировались . Har'a liphatsa tsa lefutso tsa 35,170 tse hlahlobiloeng, 2917 ha lia ka tsa hlahisoa 'me liphatsa tsa lefutso tse setseng tsa 4785 li ne li e-na le polelo e sa utloahaleng (botlaaseng bo bolelang <5).Ka hona, palo ea liphatsa tsa lefutso e nkoang e hlalositsoe (motheo o bolela ≥ 5) nakong ea teko e ne e le 27,468 (78.1%) (Supplementary Table S4).
Ho tloha ka lekhetlo la pele, melapo eohle ea NLL e ile ea arabela ho inoculation ea C. lupini (strain Col-08) ka ho tsosolosa transcriptome (Letlapa la 1), leha ho le joalo, ho ile ha hlokomeloa phapang e khōlō pakeng tsa mela.Ka hona, mohala oa ho hanyetsa 83A: 476 (e jereng liphatsa tsa lefutso tsa Lanr1) o bontšitse phetoho e kholo ea transcriptome ka lekhetlo la pele (6 hpi) ka keketseho ea 31-69 ea palo ea liphatsa tsa lefutso tse ka thōko ho bapisoa le lintlha tse ling tsa nako ka nako ena.Ho phaella moo, tlhōrō ena e ne e le ea nakoana, kaha polelo ea liphatsa tsa lefutso tse seng kae feela e ile ea lula e fetotsoe haholo ka nako ea bobeli (12 hpi).Hoa thahasellisa hore, Boregine, eo hape e bontšitseng boemo bo phahameng ba khanyetso tekong ea graft, ha ea ka ea kena ho reprogramming e kholo joalo ea transcriptional nakong ea teko.Leha ho le joalo, palo ea liphatsa tsa lefutso tse fapaneng (DEG) e ne e tšoana bakeng sa Boregine le 83A: 476 ho 12 HPI.Ka bobeli Mandelup le baahi ba 22660 ba bontšitse litlhōrō tsa DEG ka nako ea ho qetela (48 l / s), e bontšang ho lieha ho lekanyelitsoeng ho likarabo tsa tšireletso.
Hobane 83A: 476 e ile ea e-ba le reprogramming e kholo ea transcriptome ho arabela C. lupini ho 6 HPI ha e bapisoa le mela e meng eohle, ~ 91% ea li-DEG tse hlokometsoeng ka nako ena li ne li khethiloe ka ho toba (setšoantšo sa 1).Leha ho le joalo, ho bile le ho kopana ho hong ho likarabelo tsa pele lipakeng tsa mela ea boithuto, joalo ka 68.5%, 50.9%, le 52.6% DEG ho Boregine, Mandelup, le palo ea batho ba 22660, ka ho latellana, e fetelletseng le e fumanoang ho 83A: 476 ka lintlha tse itseng ka nako.Leha ho le joalo, li-DEG tsena li ne li e-na le karoloana e nyenyane feela (0.97-1.70%) ea li-DEG tsohle tse fumanoeng hona joale ho sebelisoa 83A: 476.Ho phaella moo, li-DEG tsa 11 tse tsoang meleng eohle li ne li lumellana ka nako ena (Supplementary Tables S4-S6), ho kenyelletsa le likarolo tse tloaelehileng tsa likarabo tsa tšireletso ea limela: protheine ea ho fetisa lipid (TanjilG_32225), endoglucan-1,3-β-glucoside enzyme (TanjilG_23384), tse peli tsa khatello ea kelello-inducible Tag52_3 protheine ea TanjilG5-3 (TanjilG531) 31), protheine ea motheo ea latex (TanjilG_32352), le liprotheine tse peli tse nang le glycine e ngata ea lerako la lisele (TanjilG_19701 le TanjilG_19702).Ho ne ho boetse ho e-na le ho kopana ho batlang ho phahame ho likarabo tsa transcriptome pakeng tsa 83A: 476 le Boregine ho 24 HPI (kakaretso ea 16-38% DEG) le pakeng tsa Mandelup le Population 22660 ho 48 HPI (kakaretso 14-20% DEG).
Sets'oants'o sa Venn se bonts'ang palo ea liphatsa tsa lefutso tse hlahisoang ka mokhoa o fapaneng (DEG) meleng e moqotetsane ea lupine (NLL) e kentsoeng ka Colletotrichum lupini (strain Col-08 e fumanoeng masimong a lupine Wierzhenice, Poland, 1999).Mela ea NLL e hlahlobiloeng e ne e le: 83A: 476 (e hanyetsanang, e jereng Lanr1 allele), Boregine (e hanyetsanang, liphatsa tsa lefutso tse sa tsejoeng), Mandelup (e hanyetsanang ka mokhoa o itekanetseng, e jereng AnMan allele) le baahi ba 22660 (ba hlaselehang habonolo haholo).Kgutsufatso ya hpi e emetse dihora tse kamora ho entoa.Litekanyetso tsa Zero li tlositsoe ho nolofatsa graph.
Sehlopha sa liphatsa tsa lefutso tse fetelletseng ho 6 hpi se ile sa hlahlobjoa bakeng sa ho ba teng ha libaka tsa liphatsa tsa lefutso tsa R (Supplementary Table S7).Phuputso ena e senotse ho kenngoa ha transcriptome ea liphatsa tsa lefutso tsa khale tsa ho hanyetsa mafu tse nang le libaka tsa NBS-LRR feela ho 83A: 476.Sehlopha sena se ne se e-na le mofuta o le mong oa TIR-NBS-LRR (tanjilg_05042), liphatsa tsa lefutso tse hlano tsa CC-NBS-LRR (tanjilg_06165, tanjilg_06162, tanjilg_22773, tanjilg_22640, le tanjilg_16162R), Four Tanjilg_16162RB, Four Tanjilg_16162-BSR RRE (tanjilg_16162) hammoho le Four NBS-Lrr (tanjilg_16162) le Four NBS-LRR (TANJILG_16162).Liphatsa tsa lefutso tsena kaofela li na le likarolo tsa molao tse hlophisitsoeng ka tatellano e bolokiloeng.Ho phaella ho liphatsa tsa lefutso tsa NBS-LRR, li-RLL kinase tse 'maloa li ile tsa sebelisoa ho 6 hpi, e leng e le 'ngoe Boregine (TanjilG_19877), tse peli Mandelup (TanjilG_07141 le TanjilG_19877) le ka baahi ba 22660 (TanjilG_06877) le tse peli tse Tanjil 3: 37014 le TanjilG_09014 6.
Liphatsa tsa lefutso tse nang le polelo e fetotsoeng haholo ho arabela ho ente ka C. lupini (strain Col-08) li ne li le tlas'a tlhahlobo ea ntlafatso ea Gene Ontology (GO) (Supplementary Table S8). Nako ea ts'ebetso ea likokoana-hloko e atisang ho hlalosoa ka ho fetisisa e ne e le "GO: 0006952 karabo ea tšireletso" e hlahang ho 6 ho tsoa ho 16 (nako × line) e nang le bohlokoa bo phahameng (P boleng <0.001) (Setšoantšo sa 2). Nako ea ts'ebetso ea likokoana-hloko e atisang ho hlalosoa ka ho fetisisa e ne e le "GO: 0006952 karabo ea tšireletso" e hlahang ho 6 ho tsoa ho 16 (nako × line) e nang le bohlokoa bo phahameng (P boleng <0.001) (Setšoantšo sa 2). Наиболее часто чрезмерно представленным термином биологического процесса был "GO: 0006952 защитный отвенным термином биологического процесса был "GO: 0006952 защитный ответ», который песни 6 × 6 комбинаций с высокой значимостью (значение P <0,001) (рис. 2). Nako ea ts'ebetso ea likokoana-hloko e atisang ho etsoa hangata e ne e le 'GO: 0006952 karabo ea ts'ireletso', e hlahang ho 6 ea 16 (nako × lineage) e nang le bohlokoa bo phahameng (P boleng <0.001) (Setšoantšo sa 2).最常被过度代表的生物过程术语是“GO:0006952 防御反应”，它出现在16 个)着性 (P 值 <0.001) (图2) Nako e emelang ka ho fetisisa ea ts'ebetso ea likokoana-hloko ke "GO: 0006952 karabo ea tšireletso", e hlahang ho 6 ea 16 (时间× 线) metsoako, e nang le bohlokoa bo phahameng (P boleng <0.001) (图2) . Наиболее часто чрезмерно представленным термином биологического процесса был "GO: 0006952 Defense Response", который появлялся 10 × с высокой значимостью (значение P <0,001) (рис. 2). Nako ea ts'ebetso ea likokoana-hloko e atisang ho etsoa hangata e ne e le 'GO: 0006952 Tšireletso Response', e hlahileng ka 6 ho tsoa ho 16 metsoako (nako × line) e nang le bohlokoa bo phahameng (P boleng <0.001) (Setšoantšo sa 2).Lentsoe lena le ne le emeloa ho feta linako tse peli ho 83A: 476 le Boregine (6 le 24 hpi) le ka nako e 'ngoe Mandelup le Population 22660 (12 le 6 hpi, ka ho latellana).Sena ke sephetho se lebelletsoeng, se totobatsang karabelo ea antifungal ea mela e hanyetsanang.Ho phaella moo, 83A: 476 e ile ea arabela C. lupini ka ho potlakisa liphatsa tsa lefutso tse amanang le ho phatloha ha oxidative e emeloang ke lentsoe "GO: 0055114 redox process", e bontšang karabo e itseng ea tšireletso, ha Boregine e senola likarabo tse khethehileng tsa tšireletso, tse amanang le lentsoe 'GO'.:0006950 Karabelo ea Khatello”.Baahi ba 22660 ba ile ba kenya tšebetsong karabelo e otlolohileng ea khanyetso e kenyelletsang metabolites ea bobeli, e totobatsang palo e feteletseng ea mantsoe "GO: 0016104 Ts'ebetso ea triterpene biosynthesis" le "GO:0006722 Ts'ebetso ea triterpene metabolism" (mantsoe ka bobeli ke a sete e tšoanang ea liphatsa tsa lefutso), ho nahanoa ka sephetho sa ntlafatso ea population le tlhahlobo ea GO. 2660. Ho feta moo, karabelo ea pele 83A:476 (6 hpi) le karabelo e liehang ho etsoa Mandelup le Population 22660 li kenyelletsa lentsoe GO:0015979 'photosynthesis' le lits'ebetso tse ling tse amanang le baeloji.
Mantsoe a bioprocess gene ontology a khethiloeng tlhalosong ea liphatsa tsa lefutso tse hlalositsoeng ka mokhoa o fapaneng nakong ea likarabo tsa transcriptome tsa lupine e nang le lekhasi le moqotetsane (NLL) e kentsoeng ka koatsi lupine (Col-08 strain e fumanoeng masimong a lupine Wierzhenice, Poland, ka 1999) a feteletsoa haholo.Mela ea NLL e hlahlobiloeng e ne e le: 83A: 476 (e hanyetsanang, e jereng homozygous Lanr1 allele), Boregine (e hanyetsanang, e sa tsejoeng ea liphatsa tsa lefutso), Mandelup (e hanyetsanang ka mokhoa o itekanetseng, e jereng homozygous AnMan allele) le baahi ba 22660 (ba hlaselehang habonolo).
Hobane thuto ena e ne e reretsoe ho khetholla liphatsa tsa lefutso tse tlatsetsang ho hanyetsa anthracnose, liphatsa tsa lefutso tse abetsoeng mantsoe a GO "GO: 0006952 Defensive responses" le "GO: 0055114 Redox processs" li ile tsa hlahlojoa ka ho khaola ho tloha ha motheo o bolela ≥ 30 bonyane ka mola o le mong.× ntlha ka nako e kopanyang boleng ba lipalo-palo ba log2 (phetoho ea menahano).Palo ea liphatsa tsa lefutso tse finyellang litekanyetso tsena e ne e le 65 bakeng sa GO:0006952 le 524 bakeng sa GO:0055114.
83A: 476 e senotse litlhōrō tse peli tsa DEG tse hlalositsoeng ka lentsoe GO: 0006952, ea pele ho liphatsa tsa lefutso tse 6 ka inch (liphatsa tsa lefutso tse 64, melao ea holimo le e tlaase) 'me ea bobeli e le liphatsa tsa lefutso tse 24 ka inch (liphatsa tsa lefutso tse 15, ho ea holimo feela).Boregine o boetse a bonts'a hore GO: 0006952 e fihlile sehlohlolong ka nako e ts'oanang, empa ka DEG e nyane (11 le 8) le ts'ebetso e ikhethileng.Mandeloop e bonts'itse litlhoro tse peli tsa GO: 0006952 ho 12 le 48 HPI, ka bobeli e nkile liphatsa tsa lefutso tse 12 (ea pele e nang le liphatsa tsa lefutso tse sebetsang, 'me ea bobeli e na le liphatsa tsa lefutso tse hatellang feela), athe palo ea batho ba 22660 ho 6 HPI (liphatsa tsa lefutso tse 13) e ne e e-na le tšusumetso e kholo ea tlhoro ea keketseho.taolo.Hoa lokela ho hlokomeloa hore 96.4% ea GO: 0006952 DEG litlhōrōng tsena e ne e e-na le mofuta o tšoanang oa karabelo (holimo kapa tlase), e bontšang ho kopana ho hoholo ha likarabo tsa tšireletso ho sa tsotellehe phapang ea palo ea liphatsa tsa lefutso tse amehang.Sehlopha se seholo ka ho fetisisa sa tatelano e amanang le lentsoe GO:0006952 e kenyelletsa Protein 22 (SAM22-like) ea Starvation Stress-Associated Message, e leng ea sehlopha sa 10 sa protheine e amanang le pathogenesis (PR-10) protein clade le core protein latex.protheine e tšoanang (MLP-like)) protheine (setšoantšo sa 3).Lihlopha tsena tse peli li ne li fapane ka sebopeho sa polelo le tataiso ea karabo.Liphatsa tsa lefutso tse kenyelletsang liprotheine tse kang tsa SAM22 li bonts'itse ts'ebetso e tsitsitseng le ea bohlokoa ka nako ea pele (6 kapa 12 hpi) 'me ka kakaretso e ne e sa arabele qetellong ea teko (48 hpi), ha liprotheine tse kang MLP li bontšitse ho sebelisana ho 6 hpi.hpi.83A: 476 le Mandelup ho 48 hp / in, hoo e batlang e le lintlha tse ling tsohle tsa data li ne li sa arabele.Ho feta moo, liphapang tsa liphatlalatso tsa liphatsa tsa lefutso tsa SAM22 tse kang protheine li ile tsa latela ho bonahala ho feto-fetoha ha ho hanyetsa anthracnose, kaha mela e mengata e hanyetsanang e ne e e-na le nako e ngata e susumetsang liphatsa tsa lefutso ho feta liphatsa tsa lefutso tse hlaselehang habonolo.Mofuta o mong oa LlR18A/B-joaloka PR-10 o bontšitse mokhoa o tšoanang oa polelo ho mofuta oa protheine o kang oa SAM22.
Likarolo tse kholo tsa poleloana ea ts'ebetso ea baeloji "GO:0006952 Defense Response" le lipaterone tsa polelo ea liphatsa tsa lefutso tsa Lanr1 le AnMan alleles li ile tsa khetholloa.Sekala sa Log2 se emela litekanyetso tsa log2 (phetoho) lipakeng tsa ente (Colletotrichum lupini, strain Col-08, e fumanoeng masimong a lupine, Wizhenica, Poland, 1999) le limela tsa taolo (sham-inoculated) ka nako e le 'ngoe.Ho ile ha hlahlobjoa mela e latelang ea letlapa le lesesaane la lupine: 83A:476 (e hanyetsanang, e jereng homozygous Lanr1 allele), Boregine (e hanyetsanang, liphatsa tsa lefutso tse sa tsejoeng), Mandelup (e hanyetsanang ka mokhoa o itekanetseng, e jereng homozygous AnMan allele), le Baahi ba 22660 (ba hlaselehang habonolo).
Ho phaella moo, litlhaloso tsa lipolelo tsa mefuta ea mokhethoa oa RNA-seq Lanr1 (TanjilG_05042) le AnMan (TanjilG_12861) li ile tsa hlahlojoa (setšoantšo sa 3).Mofuta oa TanjilG_05042 o bontšitse karabelo e kholo (ts'ebetsong) ho 83A: 476 feela ka lekhetlo la pele (6 hpi), ha TanjilG_12861 e ne e le ea bohlokoa ho Mandeloop ka linako tse peli feela: 6 hpi (taolo e tlase) le 24 hpi (6 hpi).Ka.).fetohang)).
Liphatsa tsa lefutso tse feteletseng ka ho fetisisa ka nako ea GO: 0055114 "ts'ebetso ea redox" e ne e le liphatsa tsa lefutso tse kenyang liprotheine tsa cytochrome P450 le peroxidase (setšoantšo sa 4).Bakeng sa disampole tse arohaneng le 83A: 476 ho 6 HPI, litekanyetso tse phahameng kapa tse tlase tsa log2 (phetoho e mene) (bakeng sa 86.6% ea liphatsa tsa lefutso) hangata li ne li bonoa lipakeng tsa limela tse kentsoeng le tse laolang, tse totobatsang karabelo e phahameng ea genotype ena ho enteng thobalano.83A: 476 e bontšitse GO ea bohlokoa ka ho fetisisa: 0055114 DEG ho 6 hpi (liphatsa tsa lefutso tse 503), ha mela e meng e le 48 hpi (Boregine, liphatsa tsa lefutso tsa 31; Mandelup, 85 liphatsa tsa lefutso; le Baahi ba 22660, 78 liphatsa tsa lefutso)).Liphatsa tsa lefutso tse ngata tsa lelapa la GO: 0055114, mefuta e 'meli ea likarabo ho ente (ts'ebetso le ho thibela) e ile ea hlokomeloa.Hoa thahasellisa hore ho fihlela ho 97.6% ea li-DEGs tse khethiloeng bakeng sa lentsoe GO: 0055114 ho Mandelupe ho 48 hp Litlhokomeliso tsena li fana ka maikutlo a hore, ho sa tsotellehe tekanyo e nyenyane haholo (ke hore, palo ea liphatsa tsa lefutso tsa redox tse fetotsoeng, 85 khahlanong le 503), mokhoa oa ho lieha ha likarabo tsa transcriptome ho 6 Arabano ea pele ea 6 aracnoup e tšoanang le ea 6 A8.Ho Boregine le Population 22660, kopano ena e tlase ho 51.6% le 75.6%, ka ho latellana.
Mekhoa ea polelo ea likarolo tse ka sehloohong tsa nako ea ts'ebetso ea likokoana-hloko "GO: 0055114 Redox process" e senotsoe.Sekala sa Log2 se emela litekanyetso tsa log2 (phetoho) lipakeng tsa ente (Colletotrichum lupini, strain Col-08, e fumanoeng masimong a lupine, Wizhenica, Poland, 1999) le limela tsa taolo (sham-inoculated) ka nako e le 'ngoe.Ho ile ha hlahlobjoa mela e latelang ea letlapa le lesesaane la lupine: 83A:476 (e hanyetsanang, e jereng homozygous Lanr1 allele), Boregine (e hanyetsanang, liphatsa tsa lefutso tse sa tsejoeng), Mandelup (e hanyetsanang ka mokhoa o itekanetseng, e jereng homozygous AnMan allele), le Baahi ba 22660 (ba hlaselehang habonolo).
83A: 476 Likarabo tsa Transcriptomic ho inoculation le C. lupini (strain Col-08) e boetse e kenyelletsa ho khutsisa ho hokahaneng ha liphatsa tsa lefutso tse boleloang ke lentsoe GO: 0015979 "photosynthesis" le mekhoa e meng e amanang le likokoana-hloko (FIG. 5).Sehlopha sena sa GO: 0015979 DEG se ne se na le liphatsa tsa lefutso tsa 105 tse ileng tsa hatelloa haholo ho 6 hpi ho 83A: 476.Sehlopheng sena, liphatsa tsa lefutso tse 37 le tsona li ile tsa theoloa Mandelup ho 48 HPI le 35 ka nako e le 'ngoe ho baahi ba 22660, ho kenyeletsoa 19 DEGs e tloaelehileng ho mefuta ka bobeli.Ha ho li-DEG tse amanang le nako ea GO: 0015979 li ile tsa sebelisoa haholo motsoako ofe kapa ofe (line x nako).
Mekhoa ea polelo ea likarolo tse ka sehloohong tsa nako ea ts'ebetso ea likokoana-hloko "GO: 0015979 Photosynthesis" e senotsoe.Sekala sa Log2 se emela litekanyetso tsa log2 (phetoho) lipakeng tsa ente (Colletotrichum lupini, strain Col-08, e fumanoeng masimong a lupine, Wizhenica, Poland, 1999) le limela tsa taolo (sham-inoculated) ka nako e le 'ngoe.Ho ile ha hlahlobjoa mela e latelang ea letlapa le lesesaane la lupine: 83A:476 (e hanyetsanang, e jereng homozygous Lanr1 allele), Boregine (e hanyetsanang, liphatsa tsa lefutso tse sa tsejoeng), Mandelup (e hanyetsanang ka mokhoa o itekanetseng, e jereng homozygous AnMan allele), le Baahi ba 22660 (ba hlaselehang habonolo).
Ho ipapisitsoe le liphetho tsa tlhahlobo ea polelo e fapaneng le eo ho ka etsahalang hore e kentse letsoho likarabelong tsa ts'ireletso khahlano le li-fungus tsa pathogenic, sehlopha sena sa liphatsa tsa lefutso tse supileng se ile sa khethoa bakeng sa quantification of expression profiles by real-time PCR (Supplementary Table S9).
The putative protein gene TanjilG_10657 e khothalelitsoe haholo meleng eohle e ithutoang le lintlha tsa nako ha li bapisoa le limela tsa taolo (Tsela ea Tlatsetso S10, S11).Ho feta moo, boemo ba polelo ea TanjilG_10657 bo bontšitse mokhoa o ntseng o eketseha nakong ea liteko bakeng sa mela eohle.Baahi ba 22660 ba bontšitse kutloisiso e phahameng ka ho fetisisa ea TanjilG_10657 ho ente e nang le ts'ebetso ea 114-fold le boemo bo phahameng ka ho fetisisa ba polelo (4.4 ± 0.4) ho 24 HPI (Fig. 6a).The PR10 LlR18A protheine gene TanjilG_27015 e boetse e bontšitse ts'ebetso ho pholletsa le mela eohle le lintlha tsa nako, ka bohlokoa ba lipalo-palo libakeng tse ngata tsa data (setšoantšo sa 6b).Ho ts'oana le TanjilG_10657, boemo bo phahameng ka ho fetisisa ba polelo ea TanjilG_27015 bo ile ba bonoa ho batho ba 22660 ba entetsoeng ho 24 HPI (19.5 ± 2.4).Asiti ea endochitinase gene TanjilG_04706 e ne e laoloa haholo meleng eohle le ka linako tsohle ntle le Boregine 6 hpi (setšoantšo sa 6c).E ile ea susumetsoa ka matla ka lekhetlo la pele (6 HPI) ho 83A: 476 (ka linako tse 10.5) 'me e eketseha ka mokhoa o itekanetseng meleng e meng (ka linako tse 6.6-7.5).Nakong ea teko, polelo ea TanjilG_04706 e ile ea lula e le maemong a tšoanang ho 83A: 476 le Boregine, ha Mandelup le Population 22660 e eketsehile haholo, e fihla ho litekanyetso tse phahameng (5.9 ± 1.5 le 6.2 ± 1.5, ka ho latellana).Mofuta oa endoglucan-1,3-β-glucosidase-joaloka TanjilG_23384 o bontšitse ts'ebetso e phahameng ka linako tse peli tsa pele (6 le 12 hpi) meleng eohle ntle le baahi ba 22660 (setšoantšo sa 6d).Maemo a phahameng ka ho fetesisa a amanang le TanjilG_23384 a ile a bonoa lekhetlong la bobeli (12 hpi) Mandelup (2.7 ± 0.3) le 83A:476 (1.5 ± 0.1).Ho 24 HPI, polelo ea TanjilG_23384 e ne e batla e le tlase meleng eohle e ithutoang (ho tloha 0.04 ± 0.009 ho ea 0.44 ± 0.12).
Tlhaloso ea liphatsa tsa lefutso tse khethiloeng (ag) tse senotsoeng ke palo ea PCR.Linomoro 6, 12 le 24 li emela lihora tse ngata ka mor'a ho entoa.Liphatsa tsa lefutso tsa LanDExH7 le LanTUB6 li ne li sebelisetsoa ho tloaeleha 'me LanTUB6 e ne e sebelisetsoa ho lekanya lipakeng tsa lihlopha.Makoko a liphoso a emela ho kheloha ho tloaelehileng ho ipapisitseng le likopi tse tharo tsa baeloji, e 'ngoe le e 'ngoe ea tsona e leng karolelano ea likopi tse tharo tsa tekheniki. Bohlokoa ba lipalo-palo ba ho se tšoane ha maemo a polelo pakeng tsa ente (Colletotrichum lupini, strain Col-08, e fumanoeng ka 1999 ho tloha tšimong ea lupine Wierzenica, Poland) le limela tsa taolo (mock-inoculated) li tšoailoe ka holimo ho lintlha tsa data (*P boleng <0.05, **P boleng ≤0 0.0 P). Bohlokoa ba lipalo-palo ba ho se tšoane ha maemo a polelo pakeng tsa ente (Colletotrichum lupini, strain Col-08, e fumanoeng ka 1999 ho tloha tšimong ea lupine Wierzenica, Poland) le limela tsa taolo (mock-inoculated) li tšoailoe ka holimo ho lintlha tsa data (*P boleng <0.05, **P boleng ≤0 0.0 P). Статистическая значимость различий в уровнях экспрессии между инокулированными (Colletotrichum lupini, штам Col-08, уровнях экспрессии между инокулированными) а) и контрольными (ложно инокулированными) растениями отмечена над точками данных (*значение P < 0,05, **значение P ≤ 0,01, *** значение 0, 0. Liphapang tsa lipalo-palo tsa maemo a polelo lipakeng tsa ente (Colletotrichum lupini, strain Col-08, e fumanoeng ka 1999 ho tloha tšimong ea lupine Wierzhenice, Poland) le limela tsa taolo (sham-inoculated) li hlokometsoe ka holimo ho lintlha tsa data (*P boleng <0.05, **P-boleng ≤ 0.0 01 ≤ ≤ 0 01 ≤ 0.0 01 ≤ 0.01 ≤ 0.01 ≤ 0.0 01.接种（Colletotrichum lupini，Col-08株，1999年从波兰Wierzenica的羽扇豆田获得）和对照（模拟接种种）植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 （colletorichum lupini ， color-08 株 ， 1999 年 波兰 波兰 wierzenica 的羽扇 获得） 和对照 鼈接种 对照 鼈接秗 波种的 统计学 显着性 标记 数据点 上方*p 值 <0.05, **P ≤ 0.01, ***P ≤ 0.001). Статистически значимые различия в уровнях экспрессии между инокулированными (Colletotrichum lupini, штамм Col-08, полученный с полей люпи, 9 10 г. Liphapang tsa lipalo-palo lipakeng tsa maemo a polelo pakeng tsa ente (Colletotrichum lupini, strain Col-08, e fumanoeng masimong a lupine Verzhenice, Poland, ka 1999) le limela tsa taolo (sham-inoculated) li hlokometsoe ka holimo ho lintlha tsa data (*P boleng <0.05 , **P-boleng ≤ 0-10 ≤ 0-11 ≤ 0 ≤ 0.0 1).Mela ea NLL e hlahlobiloeng e ne e le: 83A: 476 (e hanyetsanang, e jereng homozygous Lanr1 allele), Mandelup (e hanyetsanang ka mokhoa o itekanetseng, e jereng homozygous AnMan allele), Boregine (e hanyetsanang, e sa tsejoeng ea liphatsa tsa lefutso) le baahi ba 22660 (ba hlaselehang habonolo).
Mofuta oa mokhethoa oa TanjilG_05042 sebakeng sa Lanr1 o bontšitse mokhoa o fapaneng haholo oa polelo ho tsoa ho liprofaele tse fumanoeng ho tsoa lithutong tsa RNA-seq (setšoantšo sa 6e).Ts'ebetso e kholo ea lefutso lena e ile ea bonoa Mandelup le palo ea batho ba 22660 (ho fihlela makhetlo a 39.7 le 11.7, ka ho latellana), e leng se ileng sa fella ka maemo a phahameng a polelo (ho fihla ho 1.4 ± 0.14 le 7.2 ± 1.3, ka ho latellana).83A: 476 e boetse e senotse ho phahama ha mofuta oa TanjilG_05042 (ho fihla ho makhetlo a 3.8), leha ho le joalo, maemo a lekanyelitsoeng a polelo a fihletsoeng (0.044 ± 0.002) a ne a le tlase ka makhetlo a 30 ho feta a bonoang Mandelup le batho ba 22660.e hlahlobiloeng ke qPCR e bontšitse phapang e kholo ea maemo a polelo pakeng tsa genotypes ka mefuta e fapaneng ea mock-vaccinated (taolo), ho fihlela phapang ea 58 pakeng tsa baahi ba 22660 le 83A: 476, hammoho le pakeng tsa batho ba 22660 le 22660. Phapang e 'meli e ile ea finyelloa pakeng tsa Boregine le Mandalup.
Lefutso la mokhethoa sebakeng sa AnMan, TanjilG_12861, le ile la kenngoa tšebetsong ka lebaka la ente ka 83A:476 le Mandelup, le ne le sa nke lehlakore sechabeng sa 22660, 'me le ile la fokotsoa Boregine (Setšoantšo sa 6f).Polelo e lekanyelitsoeng ea mofuta oa TanjilG_12861 e ne e le eona e phahameng ka ho fetesisa ho 83A: 476 (0.14±0.01).Sehlopha sa 17.4 kDa sehlopha sa I sa mocheso oa liprotheine tsa mofuta oa TanjilG_05080 HSP17.4 se bontšitse maemo a tlase a lekanyelitsoeng ho mefuta eohle e ithutoang le lintlha tsa nako (setšoantšo sa 6g).Theko e phahameng ka ho fetisisa e ile ea hlokomeloa ho 24 HPI ho baahi ba 22660 (0.14 ± 0.02, keketseho ea makhetlo a robeli karabong ea ente).
Papiso ea liphatlalatso tsa liphatsa tsa lefutso (setšoantšo sa 7) e senotse kamano e phahameng pakeng tsa TanjilG_10657 le liphatsa tse ling tse 'nè: TanjilG_27015 (r = 0.89), TanjilG_05080 (r = 0.85), TanjilG_05042 (r = 0.80G = 0.80) = 0.80 (r = 0.80) = 0.80 G_9).Liphetho tse joalo li ka bonts'a taolo e kopanetsoeng ea liphatsa tsa lefutso nakong ea likarabo tsa ts'ireletso.Liphatsa tsa lefutso tsa TanjilG_12861 le TanjilG_23384 li bonts'itse liprofaele tse fapaneng tsa polelo tse nang le boleng bo tlase ba khokahano ea Pearson (ho tloha ho 0.08 ho isa ho 0.43 le -0.19 ho isa ho 0.28, ka ho latellana) ha ho bapisoa le mefuta e meng.
Likamano lipakeng tsa li-profaele tsa liphatsa tsa lefutso li ile tsa bonoa ho sebelisoa palo ea PCR.Ho ile ha hlahlobjoa mela e latelang ea letlapa le lesesaane la lupine: 83A:476 (e hanyetsanang, e jereng homozygous Lanr1 allele), Mandelup (e hanyetsanang ka mokhoa o itekanetseng, e jereng homozygous AnMan allele), Boregine (e hanyetsanang, semelo sa lefutso se sa tsejoeng), le Baahi ba 22660 (ba hlaselehang habonolo).Ho ile ha baloa lintlha tse tharo (lihora tse 6, 12 le 24 ka mor'a ho entoa), ho kenyeletsoa ente (Colletotrichum lupini, strain Col-08, e fumanoeng masimong a lupine a Wierzhenice, Poland, ka 1999) le limela tsa taolo (sham-inoculated).Sekala se bontša boleng ba coefficient ea khokahano ea Pearson.
Ho itšetlehile ka boitsebiso bo fumanoeng ho 6 horsepower ka inch, WGCNA e entsoe ka 9981 DEG e khetholloang ka ho bapisa limela tse kentsoeng le tse laolang ho tsepamisa maikutlo ho likarabo tsa pele tsa tšireletso (Supplementary Table S12).Li-module tse mashome a mabeli a metso e 'meli tsa liphatsa tsa lefutso (lihlopha) li fumanoe li na le litlhaloso tse amanang (tse ntle kapa tse mpe) pakeng tsa genotypes le mefuta e fapaneng ea liteko. Ka karolelano, maemo a liphatsa tsa lefutso a ne a theoha ka tatellano 83A: 476 > Mandelup > Boregine > Baahi ba 22660 (ka mefuta e 'meli, leha ho le joalo, mokhoa ona o ne o le matla ho limela tsa taolo). Ka karolelano, maemo a liphatsa tsa lefutso a ne a theoha ka tatellano 83A: 476 > Mandelup > Boregine > Baahi ba 22660 (ka mefuta e 'meli, leha ho le joalo, mokhoa ona o ne o le matla ho limela tsa taolo). В среднем уровни экспрессии генов снижались в порядке 83A:476 Mandelup > Boregine > Population 22660 растений). Ka karolelano, maemo a liphatsa tsa lefutso a fokotsehile ka tatellano ea 83A: 476 > Mandelup > Boregine > Baahi ba 22660 (ka mefuta e 'meli, leha ho le joalo, mokhoa ona o ne o le matla ho limela tsa taolo).平均而言，基因表达水平按83A:476 > Mandelup > Boregine > Population 22660 的顺序下降（然而，在两种种种变中，在两种种中变变生更强.平均 而 言 , 基因 水平 按 按 83a: 476> mandelup> boregine> baahi 22660 的顺序 下降 （，在种 中 中 在 种 中 中 在 种 中 中 。 ........ ........ ........ В среднем уровни экспрессии генов снижались в ряду 83A:476 Mandelup > Boregine > Population 22660 (однако в обоих вариантах эта тенденция тенденция ний). Ka karolelano, maemo a liphatsa tsa lefutso a fokotsehile letotong la 83A: 476 > Mandelup > Boregine > Baahi ba 22660 (leha ho le joalo, ka mefuta e 'meli, mokhoa ona o ne o le matla ho limela tsa taolo).Ho entoa ho ile ha fella ka ho phahamisoa ha liphatsa tsa lefutso, haholo-holo ho li-module tsa 18, 19, 14, 6 le 1 (ka tatellano e theohang ea phello), taolo e mpe (mohlala, li-module 9 le 20) kapa ka liphello tse sa nke lehlakore (mohlala, 11, 22, 8 le 13).GO term enrichment analysis (Supplementary Table S13) e senotse "GO: 0006952 Protective responses" bakeng sa mojule o kentsoeng (18) o nang le ts'ebetso e phahameng, ho kenyeletsoa liphatsa tsa lefutso tse hlahlojoang ke qPCR (TanjilG_04706, TanjilG_23384, TanjilG_1065G difoto tse ngata haholo le TanjilG_10657) li-module tsa qalo (9).Mojule 18 concentrator (setšoantšo sa 8) e ile ea khetholloa e le mofuta oa TanjilG_26536 o kenyang protheine ea PR-10-joaloka LlR18B, 'me module 9 concentrator e ile ea tsejoa e le mofuta oa TanjilG_28955 o kenyang protheine ea photosystem II PsbQ.Mofuta oa khanyetso oa anthracnose Lanr1, TanjilG_05042, o fumanoe ho mojule oa 22 (Setšoantšo sa 9) mme o amahanngoa le mantsoe a reng "GO:0044260 Cellular macromolecular metabolic process" le "GO:0006355 Transcriptional regulation, DNA templating1G2G01 Tag.mofuta oa liphatsa tsa lefutso o kenyelletsa mocheso oa khatello ea maikutlo A-4a (HSFA4a).
Tlhahlobo ea marang-rang e lekantsoeng ea liphatsa tsa lefutso tsa li-modules tse nang le mekhoa e mengata ea likokoana-hloko "GO: 0006952 Likarabo tsa Tšireletso".Ligation e nolofalitsoe ho hlakisa mefuta e mene e hlahlobiloeng ke qPCR (TanjilG_04706, TanjilG_23384, TanjilG_10657 le TanjilG_27015).
Tlhahlobo ea marang-rang e lekantsoeng ea liphatsa tsa lefutso tsa mojule tse nang le poleloana ea "GO: 0006355: Transcriptional regulation, DNA templating" le ho jara mokhethoa oa anthracnose resistance gene Lanr1 TanjilG_05042.Ligation e nolofalitsoe ho arola mofuta oa TanjilG_05042 le mofuta oa TanjilG_01212 o bohareng.
Tlhahlobo ea ho hanyetsa anthracnose e bokelletsoeng Australia e bontšitse hore boholo ba mefuta e sa tsoa lokolloa e ne e le bonolo;Kalya, Coromup le Mandelup li hlalositsoe e le tse hanyetsanang ka mokhoa o itekanetseng, ha Wonga, Tanjil le 83A:476 li hlalositsoe e le tse hanyetsanang haholo26,27,31.e ne e e-na le allele ea ho hanyetsa e tšoanang, e khethiloeng Lanr1, 'me Coromup le Mandelup li ne li e-na le allele e fapaneng, e khethiloeng ke AnMan10, 26, 39, ha Kalya a fetisetsa allele e fapaneng., Lana2.Ho hlahlojoa ha anthracnose ho hanyetsa Jeremane ho entse hore ho khethoe mohala o hanyetsanang oa Bo7212 o nang le mokhethoa oa allele ntle le Lanr1, e khethiloeng LanrBo36.
Boithuto ba rona bo senotse maqhubu a tlase haholo (hoo e ka bang 6%) ea Lanr1 allele ho kokoana-hloko e lekiloeng.Tlhaloso ena e lumellana le liphello tsa ho hlahloba likokoana-hloko tsa Europe Bochabela ho sebelisa matšoao a Anseq3 le Anseq4, a bontšang hore Lanr1 allele e teng meleng e 'meli feela ea Belarus.Sena se fana ka maikutlo a hore Lanr1 allele ha e e-so sebelisoe haholo ke mananeo a ho ikatisa a sebakeng seo, ho fapana le Australia, moo e leng e 'ngoe ea li-allele tsa bohlokoa bakeng sa ho ikatisa ka thuso ea matšoao.Sena se ka 'na sa bakoa ke boemo bo tlaase ba khanyetso bo fanoeng ke Lanr1 allele maemong a masimo a Europe ha ho bapisoa le tlaleho ea Australia.Ho phaella moo, liphuputso tsa anthracnose libakeng tse nang le pula e ngata Australia li bontšitse hore likarabo tsa ho hanyetsa tse kopanetsoeng ke Lanr1 allele li ka 'na tsa se ke tsa sebetsa maemong a leholimo a ratang khōlo le tsoelo-pele e potlakileng ea pathogen19,42.Ha e le hantle, thutong ea hona joale, matšoao a mang a anthracnose a ne a boetse a hlokomeloa ka liphatsa tsa lefutso tse jereng Lanr1 allele, ho fana ka maikutlo a hore khanyetso e ka nyamela tlas'a maemo a nepahetseng bakeng sa nts'etsopele ea C. lupini.Ho phaella moo, litlhaloso tse fosahetseng tse fosahetseng tsa boteng ba matšoao a Anseq3 le Anseq4, tse ka bang 1 cm ho tloha sebakeng sa Lanr1, li ka khoneha 28,30,43 .
Boithuto ba rona bo bontšitse hore 83A: 476, e jereng Lanr1 allele, e ile ea arabela ho C. lupini inoculation le transcriptome reprogramming e kholo ka nako ea pele e hlahlobiloeng (6 hpi), ha e ntse e le Mandelup, e jereng AnMan allele, likarabo tsa transcriptomic li ile tsa hlokomeloa hamorao haholo.(ho tloha ho 24 ho isa ho 48 hp).Liphetoho tsena tsa nakoana likarabong tsa tšireletso li amahanngoa le ho se tšoane ha matšoao a mafu, ho totobatsa bohlokoa ba ho lemoha ha likokoana-hloko tsa pele bakeng sa karabelo e atlehileng ea ho hanyetsa.Ho tšoaetsa lisele tsa semela, li-spores tsa koatsi li tlameha ho feta methating e mengata ea nts'etsopele sebakeng sa semela, ho kenyeletsoa ho mela, ho arohana ha lisele, le ho thehoa ha appressorium.Sehlomathiso ke sebopeho se tšoaetsanoang se khomarelang bokaholimo ba moamoheli 'me se thusa ho kenella ka har'a lisele tse amohelang.Ka hona, li-spores tsa C. gloeosporioides ka pea extract li bontšitse karohano ea pele ea nucleus ka mor'a metsotso e 75-90 ea ho kenngoa, ho thehoa ha kokoana ea kokoana-hloko ka mor'a metsotso e 90-120, le khatello ka mor'a lihora tse 4 45.Mango C. gloeosporioides e bonts'itse ho mela ho fetang 40% ka mor'a lihora tse 3 tsa incubation le hoo e ka bang 20% ​​ea ho thehoa ha li-appressors ka mor'a lihora tse 4.Mofuta oa CAP20 o amanang le virulence oa C. gloeosporioides o bontšitse ts'ebetso ea mongolo ho epiphyte-forming conidia ka mor'a hore 3.5 h incubation ka boka ba avocado e nang le metsoako e phahameng ea CAP20 protheine ka mor'a 4 h 46 min.Ka mokhoa o ts'oanang, ts'ebetso ea liphatsa tsa lefutso tsa melanin biosynthesis C. trifolii e ile ea kenngoa nakong ea ho kenngoa ha lihora tse 2 ho lateloa ke ho thehoa ha appressorium ka mor'a hora ea 1.Liphuputso tsa mahlaku a makhasi li bontšitse hore fragole e kenngoeng ka C. acutatum e na le khatello ea pele ho 8 hpi, ha tamati e kenngoa ka C. coccodes e na le khatello ea pele ho 4 hpi48,49.haholo e tsamaellana le sekala sa nako sa Colletotrichum spp.ts'oaetso ea ts'oaetso.Likarabo tse potlakileng tsa tšireletso ho 83A: 476 li fana ka maikutlo a ho kenya letsoho ha ho hanyetsa limela le liphatsa tsa lefutso tsa ho itšireletsa mafung (ETI) moleng ona, ha likarabo tse liehang tsa Mandelup li tšehetsa khopolo-taba ea micro-associated molecular pattern-triggered immune (MTI) hypothesis 50. Likarabo tsa pele ho 83A: 476 le Mandelup.Ho kopana ha karolo e itseng pakeng tsa liphatsa tsa lefutso tse laoloang holimo kapa tse tlaase ka ho lieha ho arabela ho boetse ho tšehetsa khopolo ena, kaha ETI e atisa ho nkoa e le karabo e potlakileng le e ntlafetseng ea MTI e qetellang ka lefu la lisele tse hlophisitsoeng sebakeng sa tšoaetso, se tsejoang e le anaphylactic shock 51,52.
Bongata ba liphatsa tsa lefutso tse hlahisoang ke lentsoe le hlahisitsoeng haholo Gene Ontology GO:0006952 "Defense Response" ke li-homologues tse 11 tsa molaetsa oa ho itima lijo o bakoang ke khatello ea maikutlo 22 protein (e ts'oanang le SAM22) le tse supileng tse kholo tsa latex protein-likes (MLPs).-joaloka liprotheine 31, 34, 43 le 423 li bontšitse ho tšoana ho latellana.Liphatsa tsa lefutso tse kang SAM22 li bontšitse ts'ebetso e kholo e ileng ea nka nako e telele, e bontšang ho eketseha ha maemo a ho hanyetsa anthracnose (83A: 476 le Boregine).Leha ho le joalo, liphatsa tsa lefutso tse kang MLP li ile tsa theoleloa feela meleng e jereng khanyetso ea mokhethoa allele (83A:476/Lanr1 ho 6 hpi le Mandelup/AnMan ho 24 hpi).Ho lokela ho hlokomeloe hore li-homologues tse kang SAM22 kaofela li tsoa sehlopheng sa liphatsa tsa lefutso tse nkang hoo e ka bang 105 kb, ha liphatsa tsa lefutso tse kang MLP li tsoa libakeng tse arohaneng tsa genome.Ts'ebetso e hokahaneng ea liphatsa tsa lefutso tse joalo tsa SAM22 e ile ea boela ea fumanoa thutong ea rona e fetileng ea NLL ho hanyetsa Diaporthetoxica inoculation, e fana ka maikutlo a hore e ameha likarolong tse otlolohileng tsa karabo ea tšireletso.Qeto ena e boetse e tšehetsoa ke litlaleho tsa karabelo e ntle ea liphatsa tsa lefutso tse kang SAM22 ho tsoa kotsi kapa kalafo ka salicylic acid, fungal inducers, kapa hydrogen peroxide.
Liphatsa tsa lefutso tse kang MLP li bonts'itsoe hore li arabela likhatellong tse fapaneng tsa abiotic le biotic, ho kenyeletsoa mafu a fungal a baktheria, vaerase le pathogenic mefuteng e mengata ea limela55.Litaelo tsa karabelo litšebelisanong tse itseng lipakeng tsa limela le likokoana-hloko li ne li fapana ho tloha ho ho eketseha ka matla (ke hore, nakong ea tlhaselo ea k'hothone e nang le Verticillium dahliae) ho ea ho ho fokotseha haholo (ke hore, ka mor'a tšoaetso ea sefate sa apole le Alternaria spp.)56,57.Ho fokotseha ho hoholo ha liphatsa tsa lefutso tsa MLP-like 423 ho 'nile ha hlokomeloa nakong ea tšireletso ea avocado ho tšoaetso ea F. niger le nakong ea tšoaetso ea sefate sa apole Botryosphaeria berengeriana f.cn.piricola le Alternaria alternata ke li-apole pathotypes58,59.Ho feta moo, apple calli e fetisang mofuta oa MLP-joaloka 423 e ne e e-na le pontšo e tlase ea liphatsa tsa lefutso tse amanang le ho hanyetsa 'me e ne e le kotsing ea tšoaetso ea fungal59.Ho latela Fusarium oxysporum f, mofuta oa MLP-joaloka 423 le oona o ile oa hatelloa ho kokoana-hloko e tloaelehileng ea linaoa.cn.Tšoaetso ea linaoa 60.
Litho tse ling tsa lelapa la PR-10 tse khethiloeng thutong ea rona ea RNA-seq e ne e le liphatsa tsa lefutso tsa LlR18A le LlR18B ka lebaka la ho laola holimo, hammoho le liphatsa tsa lefutso (1 gene) kapa tse theohileng (3) bakeng sa protheine ea phetisetso ea lipid DIR1..Ho feta moo, WGCNA e totobatsa mofuta oa LlR18B e le setsi sa mojule ona, o kotsing e kholo ea ho entoa 'me o na le mefuta e mengata ea tšireletso.Liphatsa tsa lefutso tsa LlR18A le LlR18B li ile tsa kenngoa ka makhasi a mosehla oa lupine ka lebaka la libaktheria tsa pathogenic, hammoho le li-stems tsa NLL ka mor'a hore D. toxica inoculation, athe homologue ea raese ea liphatsa tsa lefutso tsena, RSOsPR10, e ile ea susumetsoa ka potlako ke tšoaetso ea fungal eo ho ka etsahalang hore e kenyelelitsoe jasmonic acid, 62, e sa tšoaeang 62 tsela ea DIFIDE, DIFIDEC562, DIFIDEC562. liprotheine tsa lipalangoang tsa lipid tse hlokahalang bakeng sa ho qaleha ha mokhoa o fumanoeng oa ho hanyetsa (SAR).Ka nts'etsopele ea ts'ireletso ea ts'ireletso, protheine ea DIR1 e tsamaisoa ho tloha sebakeng sa ts'oaetso ka phloem ho kenya SAR lithong tse hole.Ho khahlisang, mofuta oa TanjilG_02313 DIR1 o ile oa khothaletsoa haholo khetlong la pele meleng ea 84A:476 le Population 22660, empa khanyetso ea anthracnose e ile ea atleha feela moleng oa 84A:476.Sena se ka 'na sa bontša hore ho na le ts'ebetso e itseng ea liphatsa tsa lefutso tsa DIR1 ho NLL, kaha li-homologues tse tharo tse setseng li ile tsa arabela ho inoculation feela ka 83A: line ea 476 ho 6 hpi,' me karabo ena e ne e lebisitsoe tlaase.
Thutong ea rona, likarolo tse tloaelehileng tse lumellanang le ts'ebetso ea tlhaho e bitsoang "GO: 0055114 Redox process" e ne e le protheine ea cytochrome P450, peroxidase, linoleic acid 9S-/13S-lipoxygenase, le 1-aminocyclopropane-1-carboxylic acid oxidase.Ho feta moo, WGCNA ea rona e hlalosa HSFA4a homologue e le setsi se tsamaisang limojule tse joalo ka mokhethoa oa mofuta oa Lanr1 TanjilG_05042.HSFA4a ke karolo ea taolo e itšetlehileng ka redox ea tranelo ea nyutlelie limeleng.
Liprotheine tsa Cytochrome P450 ke li-oxidoreductases tse susumetsang NADPH le / kapa karabelo ea hydroxylation e itšetlehileng ka O2 ka metabolism ea mantlha le ea bobeli, ho kenyelletsa metabolism ea xenobiotics, hammoho le lihormone, mafura acid, sterols, likarolo tsa lebota la sele, biopolymers, le biosynthesis ea ts'ebetso ea ts'ireletso ho semela sa 60 se fokotsehileng ho P460. 10.6 log2 (fetola phetoho) ho 5.7 ka lebaka la palo e kholo ea li-homologues tse fetotsoeng (37) le liphapang tsa mekhoa ea ho arabela pakeng tsa liphatsa tsa lefutso tse itseng, tse bontšang phetoho e holimo..Ho sebelisa data ea RNA-seq feela ho hlakisa ts'ebetso ea tlhaho ea tlhaho ea liphatsa tsa lefutso tsa NLL lelapeng le leholo joalo la protheine e ka ba taba e inahaneloang haholo.Leha ho le joalo, ke habohlokoa ho hlokomela hore liphatsa tsa lefutso tse ling tsa cytochrome P450 li amahanngoa le ho eketseha ha khanyetso ho li-fungus kapa libaktheria tsa pathogenic, ho kenyelletsa le monehelo oa ho itšoara hantle69,70,71.
Li-peroxidase tsa Sehlopha sa III ke li-enzyme tsa limela tse sebetsang ka bongata tse amehang mekhoeng e mengata ea metabolic nakong ea kholo le nts'etsopele ea semela, hammoho le ho arabela khatellong ea tikoloho joalo ka letsoai, komello, matla a khanya a phahameng, le tlhaselo ea likokoana-hloko72.Li-peroxidase li ameha ho sebelisaneng ha mefuta e mengata ea limela le Anthracis, ho akarelletsa le Stylosanthes humilis le C. gloeosporioides, Lens culinaris le C. truncatum, Phaseolus vulgaris le C. lindemuthianum, Cucumis sativus le C. lagenarium73,764,75.75.Karabelo e potlakile haholo, ka linako tse ling esita le ho 4 HPI, pele fungus e kenella lesela la semela73.Mofuta oa peroxidase o ile oa boela oa arabela ho ente ea D. toxica NLL.Ho phaella mesebetsing ea bona e tloaelehileng ea ho laola ho phatloha ha oxidative kapa ho felisa khatello ea oxidative, li-peroxidase li ka kena-kenana le kholo ea likokoana-hloko ka ho etsa mekoallo ea 'mele e thehiloeng holim'a matla a lerako la lisele nakong ea lignification, subunit kapa cross-linking ea metsoako e itseng.Mosebetsi ona o ka amahanngoa le silika ho TanjilG_03329 gene e kenyang lignin-forming anion peroxidase e neng e laoloa haholo thutong ea rona ho 83A: 476 line e hanyetsanang le 6 HPI, empa eseng ka mefuta e meng le lintlha tsa nako tse sa kang tsa arabela.
9S-/13S-lipoxygenase ea linoleic acid ke mohato oa pele tseleng ea oxidative ea lipid biosynthesis78.Lihlahisoa tsa tsela ena li na le mesebetsi e mengata ea ts'ireletso ea limela, ho kenyelletsa le ho matlafatsa lerako la lisele ka ho thehoa ha li-callose le pectin deposits, le ho laola khatello ea oxidative ka tlhahiso ea mefuta ea oksijene e sebetsang79,80,81,82,83.Thutong ea hona joale, polelo ea linoleic acid 9S-/13S-lipoxygenase e fetotsoe ka mefuta eohle, empa ho batho ba nang le ts'oaetso ea 22660, ho ne ho e-na le maemo a phahameng ka linako tse fapaneng, ha a ntse a le mathateng a jereng Lanr1 le AnMan allele, e totobatsa ho fapa-fapana ha lera la oxylipinnth ho oxylipinnth reaction.
1-aminocyclopropane-1-carboxylate oxidase (ACO) homologue e ne e laoloa haholo (liphatsa tsa lefutso tsa 9) kapa e fokotsehile (liphatsa tsa lefutso tsa 2) ha e kenngoa ka lupine.Ntle le mekhelo e 'meli, likarabo tsena kaofela li etsahetse ho 6 hp.ka 83A:476.Karabelo ea enzymatic e kentsoeng ke liprotheine tsa ACO ke mohato o fokotsang sekhahla tlhahisong ea ethylene mme ka hona e laoloa haholo84.Ethylene ke hormone ea limela e bapalang likarolo tse fapaneng ho laola nts'etsopele ea limela le ho arabela maemo a abiotic le a biotic.Ho kenngoa ha mongolo oa ACO le ho kenya tšebetsong tsela ea pontšo ea ethylene ho ameha ho eketsa ho hanyetsa raese ho hemibiotrophic fungus oryzae oryzae ka ho laola tlhahiso ea mefuta e sebetsang ea oksijene le phytoalexins.Ts'ebetso e ts'oanang haholo ea ts'oaetso ea makhasi e fumanoang pakeng tsa M. oryzae le C. lupini88,89, khahlanong le morao-rao oa bohlokoa ba ho tsosolosoa ha li-homologues tsa ACO ho 83A: 476 line e tlalehiloeng thutong ena, e fetola monyetla oa ho fana ka khanyetso ho NLL anthracnose Ethylene e bontšang mohato oa bohareng litseleng tsa limolek'hule.
Thutong ea hajoale, khatello e kholo ea liphatsa tsa lefutso tse amanang le photosynthesis e ile ea bonoa ho 6 hpi ho 83A: 476 le ho 48 hpi Mandeloop le baahi ba 22660.Boemo le tsoelo-pele ea liphetoho tsena li lekana le boemo.Ho hanyetsa anthracnose ho ile ha bonoa tekong ena.Haufinyane tjena, khatello e matla le ea pele ea lingoliloeng tse amanang le photosynthesis e tlalehiloe mefuteng e mengata ea litšebelisano tsa limela-pathogen, ho kenyelletsa le libaktheria tsa pathogenic le li-fungus.Lepotlapotla (ho tloha ho 2 HPI litšebelisanong tse ling) le khatello ea lefats'e ea liphatsa tsa lefutso tse amanang le photosynthesis ho arabela tšoaetso e ka etsa hore limela li itšireletse ka lebaka la ho tsamaisoa ha mefuta ea oksijene e sebetsang le ho sebelisana ha bona le tsela ea salicylic acid ho etsa phehisano ea ho kula 90,94.
Qetellong, mekhoa ea ho arabela ts'ireletso e reriloeng bakeng sa leloko le hanyetsanang ka ho fetisisa (83A: 476) e kenyelletsa ho lemoha ka potlako pathogen ke R gene (mohlomong TIR-NBS-LRR TanjilG_05042) le allergic reaction-mediated salicylic acid le ethylene signing, e lateloa ke ho thehoa ha SAR ea nako e telele.ketso e tšehetsoa ke protheine ea DIR-1.Hoa lokela ho hlokomeloa hore nako ea biotrophic bakeng sa tšoaetso ea C. lupini e khutšoanyane haholo (hoo e ka bang matsatsi a 2), e lateloa ke necrotic growth95.Phetoho lipakeng tsa mekhahlelo ena e ka amahanngoa le necrosis le polelo ea liprotheine tse nang le ethylene-inducible tse sebetsang e le lisosa tsa karabelo ea hypersensitivity ho limela tse amohelang.Ka hona, fensetere ea nako ea ho hapa C. lupini ka katleho sethaleng sa biotrophic e moqotetsane haholo.Ho hlophisoa bocha ha liphatsa tsa lefutso tse amanang le redox le photosynthesis tse hlokometsoeng ho 83A: 476 ho 6 hpi e tsamaellana le tsoelo-pele ea fungal hyphae mme e phatlalatsa nts'etsopele ea karabelo e atlehileng ea ts'ireletso sethaleng sa biotrophic.Likarabo tsa transcriptomic tsa Mandelup le palo ea batho ba 22660 li ka lieha haholo ho tšoara fungus pele e fetohela ho kholo ea necrotic, leha ho le joalo, Mandelup e kanna ea sebetsa ho feta palo ea batho ba 22660 hobane taolo e batlang e potlakile ea protheine ea PR-10 e khothaletsa khanyetso e otlolohileng.
ETI, e tsamaisoang ke mofuta oa "R" oa canonical, e bonahala e le mokhoa o tloaelehileng oa ho hanyetsa linaoa ho anthracnose.Ka hona, ka mohlala oa legume Medicago truncatula, ho hanyetsa anthracnose ho fanoa ke mofuta oa RCT1, setho sa TIR-NBS-LRR97 semela sa mofuta oa mofuta oa R.Mofuta ona o boetse o fana ka khanyetso e pharaletseng ea anthracnose ho lesere ha le fetisetsoa ho limela tse hlaselehang habonolo.Naoang e tloaelehileng (P. vulgaris), liphatsa tsa lefutso tse fetang mashome a mabeli tsa ho hanyetsa anthracnose ho fihlela joale.Tse ling tsa liphatsa tsa lefutso tsena li fumaneha libakeng tse se nang liphatsa tsa lefutso tsa R, leha ho le joalo tse ling tse ngata li mathōkong a li-chromosome tse jereng sehlopha sa liphatsa tsa lefutso tsa NBS-LRR, ho kenyeletsoa TIR-NBS-LRRs99.Phuputso ea SSR e pharalletseng ea genome e boetse e netefalitse kamano ea mofuta oa NBS-LRR le ho hanyetsa anthracnose linaoa tse tloaelehileng.Mofuta oa "R" oa canonical o boetse o fumanoe sebakeng sa genomic o nang le sebaka se seholo sa ho hanyetsa anthracnose ka lupine e tšoeu 101.
Mosebetsi oa rona o bonts'a hore karabelo e potlakileng ea ho hanyetsa, e hlahisitsoeng qalong ea ts'oaetso ea semela (haholo-holo eseng ka morao ho 12 hpi), e sireletsa ka katleho lupine e nang le makhasi a moqotetsane ho anthracnose e bakoang ke fungus ea pathogenic Collelotrichum lupini.Re sebelisa tatellano e phahameng ea phallo, re bonts'itse lintlha tse fapaneng tsa liphatsa tsa lefutso tsa bo-anthracnose limeleng tsa NLL tse kentsoeng ke liphatsa tsa lefutso tsa Lanr1 le AnMan.Katleho ea ho itšireletsa e akarelletsa ho rala ka hloko liphatsa tsa lefutso tsa liprotheine tse amehang ho redox, photosynthesis le pathogenesis ka mor'a lihora tse seng kae ka mor'a hore semela se kopane le kokoana-hloko ea pele.Tšireletso e ts'oanang, empa e liehile ka nako, ha e sebetse haholo ho sireletsa limela maloetseng.Khanyetso ea koatsi e kentsoeng ke mofuta oa Lanr1 e tšoana le karabelo e potlakileng ea mofuta oa R (efector-triggered immune immune), athe gene ea AnMan e kanna ea fana ka karabelo e rapameng (boits'ireletso bo bakoang ke sebopeho sa molek'hule se amanang le microbe), se fana ka boemo bo itekanetseng ba botsitso.
Methapo e 215 ea NLL e neng e sebelisoa ho hlahloba matšoao a anthracnose e ne e e-na le mefuta e 74, mela e 60 e fumanoeng ka ho tšela kapa ho ikatisa, tse 5 tse fetotsoeng, le likokoana-hloko tse 76 tsa naha kapa tsa pele.Mela e ne e tsoa linaheng tse 17, haholo-holo ho tloha Poland (58), Spain (47), Jeremane (27), Australia (26), Russia (19), Belarus (7), Italy (5) le mela e meng.ho tsoa linaheng tse 10.Sehlopha se boetse se kenyelletsa mela e hanyetsanang le litšupiso: 83A:476, Tanjil, Wonga e jereng Lanr1 allele, le Mandelup e jereng AnMan allele.Mehala e ile ea fumanoa ho European Lupine Genetic Resource Database e hlokometsoeng ke Poznań Plant Breeding Ltd., Wiatrowo, Poland (Tlaleho ea Tlatsetso ea S1).
Limela li ne li lengoa tlas'a maemo a laoloang (nako ea lifoto lihora tse 16, mocheso oa 25 ° C motšehare le 18 ° C bosiu).Ho ile ha hlahlojoa likopi tse peli tsa baeloji.DNA e ne e arohane le makhasi a libeke tse tharo a sebelisa DNeasy Plant Mini Kit (Qiagen, Hilden, Germany) ho latela protocol.Boleng le mahloriso a DNA e ka thōko e ile ea hlahlojoa ka mekhoa ea spectrophotometric (NanoDrop 2000; Thermo Fisher Scientific, Waltham, MA, USA).Lesupa la AnManM1 le tšoaeang mofuta oa ho hanyetsa anthracnose AnMan (e nkiloeng ho cv. Mandelup) le matšoao Anseq3 le Anseq4 mabaleng a mofuta oa Lanr1 (e nkiloeng ho cv. Tanjil) li hlahlojoe 11,26,28.Li-Homozygote bakeng sa allele e hanyetsanang li ile tsa fuoa lintlha e le "1", e hlaselehang habonolo - e le "0", le heterozygotes - e le 0.5.
Ho ipapisitsoe le liphetho tsa tlhahlobo ea li-marker AnManM1, AnSeq3 le AnSeq4 le boteng ba lipeo bakeng sa liteko tsa ho qetela tse latelang, mela ea 50 ea NLL e khethiloe bakeng sa phenotyping ea ho hanyetsa anthracnose.Tlhahlobo e entsoe ka bobeli ka har'a sethopo se laoloang ke k'homphieutha se nang le photoperiod ea lihora tse 14 ka mocheso oa 22 ° C motšehare le 19 ° C bosiu.Lipeo lia ngolisoa (ho khaola seaparo sa peō ka lehlakoreng le leng la embryo ka lehare le bohale) pele ho jala ho thibela ho robala ha peo ka lebaka la hore seaparo sa peo se thata haholo le ho etsa bonnete ba ho mela ho tšoanang.Limela li ne li lengoa ka lipitsa (11 × 11 × 21 cm) tse nang le mobu o sa hloekang (TS-1 REC 085 Medium Basic, Klasmann-Deilmann Polska, Warsaw, Poland).Ho kenngoa ho ile ha etsoa ka mofuta oa Colletotrichum lupini Col-08, o hōlileng ka 1999 ho tloha kutung ea limela tsa lupine tse nang le makhasi a moqotetsane tse lenngoeng tšimong Verzhenítsa, Greater Poland (52° 27′ 42″ N 17° 04′ 05″ E ).Fumana sebaka.Li-isolates li ne li lengoa ka SNA medium ka 20 ° C. tlas'a khanya e ntšo bakeng sa matsatsi a 21 ho susumetsa sporulation.Libeke tse 'nè ka mor'a ho jala, ha limela li fihlile sethaleng sa makhasi a 4-6, inoculation e ne e etsoa ka ho fafatsa ka ho emisoa ha conidia ka bongata ba 0.5 x 106 conidia ka ml.Ka mor'a ho enteloa, limela li ne li bolokoa lefifing ka lihora tse 24 ka mongobo o ka bang 98% le mocheso oa 25 ° C ho nolofatsa ho mela ha conidia le ts'ebetso ea tšoaetso.Joale limela li ne li lengoa tlas'a photoperiod ea lihora tse 14 ho 22 ° C motšehare / 19 ° C bosiu le 70% ea mongobo.Palo ea lefu lena e entsoe matsatsi a 22 ka mor'a ho entoa 'me e tloha ho 0 (immune) ho ea ho 9 (e hlaselehang habonolo) ho itšetlehile ka boteng kapa ho se be teng ha liso tsa necrotic holim'a stems le makhasi.Ho phaella moo, ka mor'a ho fana ka lintlha, boima ba limela bo ne bo lekanngoa.Likamano pakeng tsa li-marker genotypes le lefu la phenotypes li ne li baloa e le lintlha tse peli tse latellanang (ho se be teng ha li-marker tsa heterozygous ka sete sa mela bakeng sa tlhahlobo ea phenotype ea ho hanyetsa anthracnose).