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Ko te Angustifolius lupine (NLL, Lupinus angustifolius L.) he tipu pīni e whakamahia ana mō te whakaputa kai me te whakapai ake i te oneone. Ko te horapa o te NLL hei hua puta noa i te ao kua kukume mai i te maha o ngā harore mate, tae atu ki te lupine anthracnose, e hua ai te mate anthracnose kino. E rua ngā alleles, ko Lanr1 me AnMan, e whakanui ake ana i te ātete, kua whakamahia i roto i te whakatipu NLL, engari kāore anō kia mōhiotia ngā tikanga ngota ngota. I roto i tēnei rangahau, i whakamahia ngā tohu Lanr1 me AnMan hei tirotiro i ngā tauira NLL Pākehā. Nā te whakamātautau i te kano kano i roto i te taiao whakahaere i whakaū te whai huatanga o ngā kaituku ātete e rua. I mahia te whakatauira ira rerekē ki runga i ngā momo ātete me ngā momo ngoikore. I honoa te ātete anthracnose ki te whakanui rawa o ngā kupu ontology ira "GO:0006952 Defense Response", "GO:0055114 Redox Process", me "GO:0015979 Photosynthesis". Hei tāpiritanga, i whakaatu te raina Lanr1(83A:476) i te tere o te whakarerekētanga o te transcriptome i muri i te werohanga, ko ētahi atu raina i whakaatu i te whakaroa o tēnei urupare mō te 42 hāora pea. Ko ngā urupare parepare e pā ana ki ngā ira TIR-NBS, CC-NBS-LRR me NBS-LRR, 10 ngā pūmua e whai wāhi ana ki te pathogenesis, ngā pūmua whakawhiti lipid, te endoglucan-1,3-β-glucosidase, ngā pūmua pakitara pūtau whai glycine, me ngā ira mai i te ara tauhohe o te hāora. Ko ngā urupare wawe ki te 83A:476, tae atu ki te pehi tupato i ngā ira e pā ana ki te photosynthesis, i rite ki te tiaki angitu i te wā o te tipu tipu o te koiora harore, e kī ana he kaiwhakaputa e whakaoho ana i te ārai mate. Ka puhoi te tauhohenga Mandeloop, me te tōia whakapae whānui.
Ko te rūpine rau-whaiti (NLL, Lupinus angustifolius L.) he witi pūmua nui e ahu mai ana i te rohe uru o te Moana-nui-a-Kiwa1,2. I tēnei wā, e whakatipuria ana hei kai mā ngā kararehe me ngā tāngata. E kiia ana hoki he paru matomato i roto i ngā pūnaha hurihuri hua nā te whakapūmau hauota e ngā huakita whakapūmau hauota me te whakapainga whānui o te hanganga oneone. Kua tere te tukanga whakawhanaungatanga o te NLL i te rautau kua hipa, ā, kei raro tonu i te pēhanga whakaputa uri teitei3,4,5,6,7,8,9,10,11,12. Nā te horapa o te whakatipu NLL, i whakawhanakehia e te raupapatanga o ngā harore mate ngā wāhi ahuwhenua hou, ā, i puta ai ngā mate hou e whakangaro ana i ngā hua. Ko te mea tino miharo ki ngā kaimahi pāmu me ngā kaiwhakatupu rūpine ko te putanga mai o te anthracnose, i puta mai i te harore mate, Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Ko te mea tino miharo ki ngā kaimahi pāmu me ngā kaiwhakatupu rūpine ko te putanga mai o te anthracnose, i puta mai i te harore mate, Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Наиболее примечательным для фермеров и селекционеров люпина было появление антракноза, селекционеров люпина было появление антракноза, вызванного памогринкого памогиного люпина (Bondar) Nirenberg, Feiler & Hagedorn13. Ko te mea tino rongonui ki ngā kaimahi pāmu me ngā kaiwhakatupu rūpine ko te putanga mai o te anthracnose i puta mai i te harore mate 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，它是由病原真菌 Lunenbergi Lungi Hagedorn13 引起的。对于羽扇豆农民和饲养者来说，最引人注目的是炭疽病的出现，它是由病原真菌lu。 Наиболее поразительным для фермеров и селекционеров люпина является появление антракноза, вызываемогего прикмтор lupini (Bondar) Nirenberg, Feiler & Hagedorn13. Ko te mea tino miharo ki ngā kaimahi pāmu me ngā kaiwhakatupu rūpine ko te putanga mai o te anthracnose i puta mai i te harore mate Colletotrichum lupini (Bondar) Nirenberg, Feiler & Hagedorn13.I ahu mai ngā pūrongo tuatahi mō te mate i Brazil me te Hononga o Amerika, ā, i puta ngā tohu noa i te tau 1912 me te tau 1929. Heoi, i muri i te 30 tau pea, i tapaina te mate ko Colletotrichum gloeosporioides (Penz.) Penz. & Sacc., te tipua nui Glomerella cingulata (Stoneman) Spauld. & Sacc., te tipua nui Glomerella cingulata (Stoneman) Spauld. & Sacc., телеоморф Glomerella cingulata (Stoneman) Spauld. & Sacc., te tino tauira o 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 i roto i te Morphology Whāia. & H. Schrenk,. & H. Schrenk, .me H. Schrenk. & H.施伦克，。 & H.施伦克，。me H. Schlenk, .Ko te whakamātautau tuatahi i ngā āhuatanga o te mate i mahia i waenganui o te rautau 20 i whakaatu i te ātete ki ngā momo NLL me te lupine kōwhai (L. luteus L.), engari ko ngā momo lupine mā katoa i whakamatautauria he tino pāngia e te mate15,16. Kua whakaatuhia e ngā rangahau ko te whanaketanga o te anthracnose e pā ana ki te pikinga o te ua (haumākū o te hau) me te pāmahana (i waenga i te 12-28°C), ka hua ake he takahi i te ātete i ngā pāmahana teitei17, 18. Inaa, ko te wā e hiahiatia ana mō ngā conidia ki te pihi me te tīmatanga o te mate, e whā ngā wā poto ake i te 24°C (4 hāora) i te 12°C (16 hāora) i raro i ngā āhuatanga haumākū teitei19. Nō reira, ko te mahana haere tonu o te ao kua arahi ki te horapa o te anthracnose. Heoi, i kitea te mate i Parani (1982) me Ukraine (1983) hei tohu mō tētahi riri e tū mai nei, engari i te āhua nei kāore i arohia e te umanga lupine i taua wā20,21. I muri i ētahi tau, ka horapa tēnei mate kino puta noa i te ao, ā, ka pāngia hoki ngā whenua nui e whakaputa ana i te lupine pērā i a Ahitereiria, Pōrana me Tiamana22,23,24. Whai muri i te pakarutanga o te mate anthracnose i waenganui o te tekau tau atu i 1990, i hua ake i te tirotiro whānui te tautuhi i ētahi kaituku ātete i roto i ngā tauira NLL19. Ko te ātete NLL ki te anthracnose e whakahaerehia ana e ngā ira matua motuhake e rua e kitea ana i roto i ngā pūtake ira tangata rerekē: Lanr1 i roto i te momo Tanjil me Wonga me AnMan i roto i te momo. Mandalay 25, 26. Ka tautokohia e ēnei ira tangata ngā tohu ngota e tautoko ana i te kōwhiringa o te ira tangata ātete i roto i ngā kaupapa whakatipu uri25,26,27,28,29,30. I whakawhitihia te raina whakatipu ātete 83A:476 e mau ana i te ira Lanr1 ki te raina mohoao ngoikore P27255 kia whiwhi ai i tētahi taupori RIL e wehea ana mō te ātete anthracnose, ā, i taea ai te tuku i te wāhi Lanr1 ki te chromosome NLL-1131, 32, 33. Mā te whakarite i ngā tohu mahere hononga mai i ngā wāhi ātete taha ki te anthracnose me te anga ira, i whakakitea e te NLL te wāhi o ngā ira e toru i runga i te chromosome kotahi (NLL-11), engari i ngā tūranga rerekē29,34,35. Heoi, nā te iti o ngā RIL me te tawhiti ira nui i waenga i ngā tohu me ngā ira e rite ana, kāore e taea te whakatau pono mō ō rātou ira kei raro. I tētahi atu taha, he uaua te whakamahi i ngā ira whakamuri i roto i ngā lupins nā te iti rawa o te kaha whakaora, e uaua ai te whakahaere ira37.
Nā te whanaketanga o te ira-pūmua kua whakangaohia e mau ana i te ira-pūmua e hiahiatia ana i roto i te āhua homozygous, pērā i te 83A:476 (Lanr1) me te Mandelup (AnMan), kua huaki te tatau ki te ako i te ātete ki te anthracnose i te aroaro o te aroaro o ngā huinga taupatupatu o ngā ira-pūmua i roto i ngā taupori mohoao. Ngā āheinga o ngā tikanga ngota. Whakatauritehia ngā urupare parepare i hangaia e ngā momoira motuhake. I arotakehia e tēnei rangahau te urupare transcriptome wawe o te NLL ki te werohanga C. lupini. Tuatahi, i tirotirohia he panui ira-pūmua NLL Pākehā kei roto ko ngā rārangi 215 mā te whakamahi i ngā tohu ngota e tohu ana i ngā ira-pūmua Lanr1 me AnMan. I muri mai ka mahia te phenotyping Anthracnose ki runga i te 50 rārangi NLL, i tīpakohia i mua mō ngā tohu ngota, i raro i ngā āhuatanga whakahaere. I runga i ēnei whakamātautau, e whā ngā rārangi he rerekē te ātete ki te anthracnose me te hanganga allelic Lanr1/AnMan i tīpakohia mō te whakatauira ira parepare rerekē mā te whakamahi i ngā huarahi tautoko e rua: te raupapatanga RNA tere-rahi me te ine PCR wā-tūturu.
I whakaatuhia e te tirotiro i tētahi huinga o te ira tangata NLL (N = 215) me ngā tohu Lanr1 (Anseq3 me Anseq4) me AnMan (Anseq4) me AnMan (AnManM1) kotahi anake te rārangi (95726, e tata ana ki Salamanca-b) e whakanui ana i te ira "ātete" mō ngā tohu katoa, i te mea i kitea e te "Aroaro o ngā ira 'ngoikore'" te ōwehenga o ngā tohu katoa i roto i te 158 rārangi (~73.5%). Tekau mā toru ngā rārangi i whakaputa i ngā ira "ātete" e rua o te tohu Lanr1, ā, e waru ngā rārangi i whakaputa i ngā ira "ātete" o Lanr1. tohu. Ko te ira "ātete" o te tohu AnMan (Ripanga Tāpiri S1). E rua ngā rārangi he heterozygous mō te tohu Anseq3 me tētahi heterozygous mō te tohu AnManM1. E 42 ngā rārangi (19.5%) i mau i ngā āhua rerekē o ngā ira Anseq3 me Anseq4, e tohu ana i te auau teitei o te whakakotahitanga i waenga i ēnei loci e rua. I whakaatuhia e ngā momo Anthracnose i raro i ngā āhuatanga whakahaere (Ripanga Tāpiri S2) te rerekētanga o te ātete o ngā momo ira i whakamatautauria, i kitea i roto i te kaha o te anthracnose. Ko ngā rerekētanga o ngā kaute toharite i ahu mai i te 1.8 (ātete āhua) ki te 6.9 (ngāwari) ā, ko ngā rerekētanga taumaha tipu i ahu mai i te 0.62 (ngāwari) ki te 4.45 g (ātete). I kitea he hononga nui i waenga i ngā uara i kitea i roto i ngā tukuruatanga e rua o te whakamātautau (0.51 mō ngā kaute kaha o te mate, P = 0.00017 me 0.61 mō te taumaha o te tipu, P < 0.0001) me waenga hoki i ēnei tawhā e rua (− 0.59 me − 0.77, P < 0.0001). I kitea he hononga nui i waenga i ngā uara i kitea i roto i ngā tukuruatanga e rua o te whakamātautau (0.51 mō ngā kaute kaha o te mate, P = 0.00017 me 0.61 mō te taumaha o te tipu, P < 0.0001) me waenga hoki i ēnei tawhā e rua (− 0.59 me − 0.77, P < 0.0001). Выявлена ​​достоверная корреляция между значениями, наблюдаемыми в двух повторностях эксперимента (0,51 для тля = 0,00017 и 0,61 для массы растения, P <0,0001), а также между этими двумя параметрами (-0,59 и -0,77, Р00 < 0,0). I kitea he hononga nui i waenga i ngā uara i kitea i roto i ngā tukuruatanga e rua o te whakamātautau (0.51 mō ngā kaute kaha o te mate, P = 0.00017 me 0.61 mō te taumaha o te tipu, P < 0.0001), me waenga hoki i ēnei tawhā e rua (- 0.59 me -0.77, P < 0.0001) 0.0001).在两次重复实验中观察到的值之间存在显着相关性（疾病严重程度评分为 =0. 0.00017，植物重量为0.61，P < 0.0001）以及这两个参数之间（- 0.59 和- 0.77，P <0.0001).在 两 次 重复 实验 中 观察 的 值 之间 存在 相关性 （疾病 严重 程度 了了 了 了 了0.51 ， p = 0.00017 ， 植物 为 为 0.61 ， p <0.0001） 以及 两 个 参数 之间 （（（（ 和 9–5. 0.59 和– 0.59 和- 0.77, P < 0.0001). Наблюдалась значительная корреляция между значениями, наблюдаемыми в двух повторностях (оценка тяжести ва, за 0,0,0,0,0,0,0,0,0,0,0,0,0,0,0,0,00,00,00,00,000 и масса растения 0,61, P <0,0001), и между этими двумя параметрами (-0,59 и -0,0001) 0,77, P <0,0001. I kitea he hononga nui i waenga i ngā uara i kitea i roto i ngā tāruarua (kaute kaha o te mate 0.51, P = 0.00017 me te taumaha tipu 0.61, P < 0.0001) ā, i waenga i ēnei tawhā e rua (-0.59 me -0 .0001) 0.77, P<0.0001. ).Ko ngā tohu noa e kitea ana i roto i ngā tipu ngoikore ko te piko me te takahuri o te kakau e rite ana ki te hanganga "piko a te hepara", ā, muri iho ko ngā whara porowhita me ngā sporozoite karaka/māwhero (Pikitia Tāpiri 1). Ko ngā momo ira o Ahitereiria e mau ana i ngā ira Lanr1 (83A:476 me Tanjil) me AnMan (Mandelup) he ātete āhua, 0.0331 me 0.0036). Ko ētahi o ngā momo ira e mau ana i ngā ira Lanr1 me/ranei AnMan "ātete" e whakaatu ana i ngā tohu o te mate.
He mea whakamīharo, i whakaatuhia e ētahi rārangi NLL kāore he ira tohu "ātete" he taumata teitei o te ātete ki te anthracnose (he rite, he teitei ake rānei i ngā momo ira Lanr1, AnMan rānei), pērā i a Boregine (uara P < 0.0001 mō ngā tawhā e rua), Bojar (uara P < 0.0001 mō te kaute me te 0.001 mō te taumaha tipu) me te Taupori B-549/79b (uara P < 0.0001 mō te kaute me te kore-hiranga mō te taumaha). He mea whakamīharo, i whakaatuhia e ētahi rārangi NLL kāore he ira tohu "ātete" he taumata teitei o te ātete ki te anthracnose (he rite, he teitei ake rānei i ngā momo ira Lanr1, AnMan rānei), pērā i a Boregine (uara P < 0.0001 mō ngā tawhā e rua), Bojar (uara P < 0.0001 mō te kaute me te 0.001 mō te taumaha tipu) me te Taupori B-549/79b (uara P < 0.0001 mō te kaute me te kore-hiranga mō te taumaha). Интересно, что несколько линий NLL, лишенных какого-либо «резистентного» маркерного аллеля, показали высокний уроский антракнозу (сопоставимый или более высокий, чем для генотипов Lanr1 или AnMan), таких как Boregine (значение P <0,0001 для проба), Boregine (значение P < 0,0001 для оценки и 0,001 для массы растения) me te популяции B-549/79b (значение P <0,0001 для оценки и незначимо для массы). He mea whakamīharo, he maha ngā momo NLL kāore he ira tohu 'ātete' i whakaatu i te taumata teitei o te ātete ki te anthracnose (he rite, he teitei ake rānei i ngā momo ira Lanr1, AnMan rānei), pērā i a Boregine (uara P < 0.0001 mō ngā tawhā e rua), Bojar (uara P < 0.0001 mō te aromatawai me te 0.001 mō te taumaha tipu) me te taupori B-549/79b (uara P < 0.0001 mō te aromatawai, ā, kāore i te hiranga mō te taumaha).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他们中文基因型相当或更高），例如Boregine（两个参数的P 值< 0.0001）、Bojar（P 值<得分为0.0001，植物重量为0.001）和种群B-549/79b（得分P 值< 0.0001，重量不显。 He mea whakamere te kite atu i ētahi pūnaha NLL kāore he tohu "antigenic" e whakaatu ana i te ātete whakapae teitei (he rite ki ngā ira Lanr1, AnMan rānei, teitei ake rānei), pērā i a Boregine (e rua ngā tawhā P < 0.0001), Bojar (uara P < 0.0001, taumaha tipu 0.001) me te momo B-549/79b (uara P < 0.0001, taumaha kāore i te hiranga). Интересно, что некоторые линии NLL, лишенные каких-либо маркерных аллелей «резистентности», показали высокие уостовички антракнозу (сравнимые или выше, чем у генотипов Lanr1 или AnMan), такие как Boregine (значение P для обоих параметров <0,0001, P 0,001), Boregine масса растения 0,001) и популяция B-549/79b (оценка P-значение <0,0001, масса незначительна). He mea whakamīharo, ko ētahi o ngā momo NLL kāore he ira tohu 'ātete' i whakaatu i ngā taumata teitei o te ātete ki te anthracnose (he rite, he teitei ake rānei i ngā momo ira Lanr1, AnMan rānei), pērā i a Boregine (uara P mō ngā tawhā e rua <0.0001), Bojar (uara-P < 0.0001, taumaha tipu 0.001) me te taupori B-549/79b (uara-P < 0.0001, taumaha kāore i te hiranga).E whakaatu ana tēnei āhuatanga i te tūponotanga o tētahi pūtake ira hou o te ātete, e whakamārama ana i te korenga o te hononga i waenga i ngā momoira tohu me ngā momoira mate (ngā uara P mai i te ~0.42 ki te ~0.98). Nō reira, i whakaatu te whakamātautau Kolmogorov-Smirnov ko ngā raraunga mō te ātete ki te anthracnose he tata te tohatoha noa mō ngā kaute (ngā uara-P 0.25 me te 0.11) me te papatipu tipu (ngā uara-P 0.47 me te 0.55), e kī ana kei te whakapae ahau he nui ake ngā ira i a Lanr1 me AnMan.
I runga i ngā hua o te tirotiro ātete ki te anthracnose, e 4 ngā momo i whiriwhiria mō te tātaritanga transcriptome: 83A:476, Boregine, Mandelup, me te Population 22660. I whakamatautauria anō ēnei momo mō te ātete ki te anthrax i roto i ngā whakamātautau werohanga mā te raupapatanga RNA, mena he rite tonu ki te whakamātautau o mua. Ko ngā uara kaute penei: Boregin (1.71 ± 1.39), 83A: 476 (2.09 ± 1.38), Mandelup (3.82 ± 1.42) me te taupori 22660 (6.11 ± 1.29).
I tutuki i te kawa Illumina NovaSeq 6000 te toharite o te 40.5 ngā takirua Mread mō ia tauira (29.7 ki te 54.4 Mreads) (Ripanga Tāpiri S3). Ko ngā kaute whakarārangi i roto i te raupapa tohutoro i ahu mai i te 75.5% ki te 88.6%. Ko te whanaungatanga toharite o ngā raraunga tatau pānui i waenga i ngā rerekētanga whakamātautau i waenga i ngā tāruarua koiora i ahu mai i te 0.812 ki te 0.997 (te toharite 0.959). Mai i ngā ira 35,170 i tātarihia, 2917 kāore i whakaatu i te whakaaturanga, ā, ko ngā ira 4785 i whakaaturia i te taumata iti noa iho (te toharite turanga < 5). Mai i ngā ira 35,170 i tātarihia, 2917 kāore i whakaatu i te whakaaturanga, ā, ko ngā ira 4785 i whakaaturia i te taumata iti noa iho (te toharite turanga < 5). Из 35 170 проанализированных генов 2917 не проявляли экспрессии, а остальные 4785 генов экспресиронвались темировались (базовое среднее <5). O ngā ira 35,170 i tātarihia, 2917 kāore i whakaatu i te whakaaturanga, ā, ko ngā ira 4785 e toe ana i whakaatuhia i te taumata iti noa iho (te toharite turanga <5).在分析的35,170 个基因中，2917 个没有表达，其他4785 个基因的表达可以忽略可以忽略不到计5）.35,170 Из 35 170 проанализированных генов 2917 не экспрессировались, a остальные 4785 генов имели незначительнсубю среднее значение <5). O ngā ira 35,170 i tātarihia, 2917 kāore i whakaatuhia, ā, ko ngā ira 4785 i toe he iti noa iho te whakaaturanga (te toharite turanga <5).Nō reira, ko te maha o ngā ira i whakaarohia kua whakaatuhia (te toharite turanga ≥ 5) i te wā o te whakamātautau he 27,468 (78.1%) (Ripanga Tāpiri S4).
Mai i te wā tuatahi, i whakautu ngā momo NLL katoa ki te werohanga o C. lupini (momo Col-08) mā te whakahou i te transcriptome (Ripanga 1), heoi, i kitea ngā rerekētanga nui i waenga i ngā momo. Nō reira, i whakaatu te momo ātete 83A:476 (e mau ana i te ira Lanr1) i te whakahounga transcriptome nui i te wā tuatahi (6 hpi) me te pikinga 31-69-ngā wā i roto i te maha o ngā ira whakarunga me ngā ira whakararo kua wehea ki te whakataurite ki ētahi atu wā i tēnei wā. Hei tāpiri, he poto noa te roa o tēnei tihi, nā te mea he torutoru noa iho ngā ira i tino rerekē i te wā tuarua (12 hpi). He mea whakamīharo, ko Boregine, i whakaatu hoki i te taumata teitei o te ātete i roto i te whakamātautau whakapiri, kāore i pāngia e te whakahounga whakawhiti nui i te wā o te whakamātautau. Heoi, ko te maha o ngā ira rerekē te whakaaturanga (DEG) he rite tonu mō Boregine me 83A:476 i te 12 HPI. I whakaatuhia e Mandelup me te taupori 22660 ngā tihi DEG i te wā whakamutunga (48 l/s), e tohu ana i te whakaroa whanaunga o ngā urupare parepare.
Nā te mea i hurihia te hōtaka transcriptome nui ki te 83A:476 hei whakautu ki te C. lupini i te 6 HPI ki te whakataurite ki ngā momo ira katoa, ~91% o ngā DEG i kitea i tēnei wā he mea motuhake ki te whakapapa (Pikitia 1). Heoi, i kitea ētahi taupatupatu i roto i ngā urupare tōmua i waenga i ngā momo ira rangahau, i te mea ko te 68.5%, 50.9%, me te 52.6% DEG i Boregine, Mandelup, me te taupori 22660, ia, i taupatupatu ki ērā i kitea i te 83A:476 i ētahi wā. Heoi, he iti noa iho te wāhanga o ēnei DEG (0.97–1.70%) o ngā DEG katoa i kitea i tēnei wā mā te whakamahi i te 83A:476. Hei tāpiritanga, e 11 ngā DEG mai i ngā momo katoa i ōrite i tēnei wā (Ngā Ripanga Tāpiri S4-S6), tae atu ki ngā wāhanga noa o ngā urupare tiaki tipu: te pūmua whakawhiti ngako (TanjilG_32225), te whākōkī endoglucan-1,3-β-glucoside (TanjilG_23384), e rua ngā pūmua ka taea te whakaoho i te ahotea pērā i a SAM22 (TanjilG_31528 me TanjilG_31531), te pūmua latex taketake (TanjilG_32352), me ētahi atu pūmua pakitara pūtau hanganga whai rawa i te glycine e rua (TanjilG_19701 me TanjilG_19702). He nui hoki te taupatupatu o ngā urupare transcriptome i waenga i te 83A:476 me Boregine i te 24 HPI (16-38% DEG katoa) me waenga i a Mandelup me te Taupori 22660 i te 48 HPI (14-20% DEG katoa).
He tūtohi Venn e whakaatu ana i te maha o ngā ira rerekē te whakaaturanga (DEG) i roto i ngā momo lupine rau-whaiti (NLL) i werohia ki te Colletotrichum lupini (momo Col-08 i tangohia mai i ngā mara lupine i Wierzhenice, Pōrana, 1999). Ko ngā momo NLL i tātarihia ko: 83A:476 (ātete, kei a ia te ira Lanr1), Boregine (ātete, kāore i te mōhiotia te papamuri ira), Mandelup (ātete āhua, kei a ia te ira AnMan) me te taupori 22660 (tino pāngia). Ko te whakarāpopototanga hpi e tohu ana i ngā hāora i muri i te werohanga. Kua tangohia ngā uara kore hei whakahaere i te kauwhata.
I tātarihia te huinga o ngā ira kua nui rawa te whakaputanga i te 6 hpi mō te aroaro o ngā rohe ira R matua (Ripanga Tāpiri S7). I whakaatuhia e tēnei rangahau te whakaurunga o ngā ira ātete mate tawhito me ngā rohe NBS-LRR i te 83A:476 anake. I titoa e tēnei huinga tētahi ira TIR-NBS-LRR (tanjilg_05042), e rima ngā ira CC-NBS-LRR (tanjilg_06165, tanjilg_06162, tanjilg_22773, tanjilg_22640, me te tanjilg_16162), me ngā NBS-LR e whā, Tanjilg_16162), me ngā NBS-LRRE e whā (tanjilg_16162) me ngā NBS-Lrr e whā (tanjilg_16162) me ngā NBS-LRR e whā (TANJILG_16162). He rohe matua kei roto i ēnei ira katoa kua whakaritea ki ngā raupapatanga kua tiakina. Haunga ngā ira rohe NBS-LRR, he maha ngā kinase RLL i whakahohehia i te 6 hpi, arā, kotahi i roto i a Boregine (TanjilG_19877), e rua i roto i a Mandelup (TanjilG_07141 me TanjilG_19877) me te taupori 22660 (TanjilG_09014 me TanjilG_10361) me te rua i roto i a 83A 27:476.
Ko ngā ira i tino rerekē te whakaaturanga i te werohanga ki te C. lupini (momo Col-08) i tukuna ki te tātaritanga whakarei ake i te Gene Ontology (GO) (Ripanga Tāpiri S8). Ko te kupu tukanga koiora i tino kitea ko te "GO:0006952 parepare urupare" i puta mai i roto i te 6 o roto i te 16 ngā huinga (wā × rārangi) me te hiranga teitei (uara P < 0.001) (Pikitia 2). Ko te kupu tukanga koiora i tino kitea ko te "GO:0006952 parepare urupare" i puta mai i roto i te 6 o roto i te 16 ngā huinga (wā × rārangi) me te hiranga teitei (uara P < 0.001) (Pikitia 2). Наиболее часто чрезмерно представленным термином биологического процесса был «GO: 0006952 защитный ответо» 6 из 16 (время × линия) комбинаций с высокой значимостью (значение P <0,001) (рис. 2). Ko te kupu tukanga koiora i tino kitea ko 'GO:0006952 urupare ārai', i puta mai i roto i te 6 o te 16 ngā huinga (wā × whakapapa) me te hiranga teitei (uara P < 0.001) (Pikitia 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（时间）一一一上一一么一上一一上个中，具有高显着性（P ‼< 0.001）（图2）。 Ko te kupu tukanga koiora tino whakaatu ko te "GO:0006952 urupare parepare", e puta ana i roto i te 6 o ngā huinga 16 (时间×线), me te hiranga teitei (uara P < 0.001) (图2). Наиболее часто чрезмерно представленным термином биологического процесса был «GO: 0006952 Defense Response», котовлыл 6 позля 6 комбинаций (время × линия) с высокой значимостью (значение P <0,001) (рис. 2). Ko te kupu tukanga koiora i tino kitea ko 'GO:0006952 Defense Response', i puta mai i roto i te 6 o roto i te 16 ngā huinga (wā × rārangi) me te hiranga teitei (uara P < 0.001) (Pikitia 2).I te 83A, i kitea nuitia tēnei kupu i ngā wā e rua: 476 me Boregine (6 me te 24 hpi) me tētahi wā i Mandelup me Population 22660 (12 me te 6 hpi). He hua tēnei e tumanakohia ana, e whakaatu ana i te urupare harore o ngā raina ātete. Hei tāpiri, i whakautu a 83A:476 ki a C. lupini mā te whakaoho tere i ngā ira e pā ana ki te pahūtanga waikura e tohuhia ana e te kupu "GO:0055114 tukanga redox", e tohu ana i tētahi urupare parepare motuhake, i te mea i whakaatu a Boregine i ngā urupare parepare motuhake, e pā ana ki te kupu 'GO'. ":0006950 Tauhohenga ahotea". I whakahohehia e te Taupori 22660 te urupare ātete whakapae e uru ana ki ngā matū matū tuarua, e whakaatu ana i te maha rawa o ngā kupu "GO:0016104 Tukanga o te koiora triterpene" me te "GO:0006722 Tukanga o te pākia o te triterpene" (nō te huinga ira kotahi ngā kupu e rua), me te whakaaro ki ngā hua o te tātaritanga whakarei kupu GO, ko te pumau o te tauhohenga Mandelup i waenganui i a Boregine me te Taupori 22660. Hei tāpiri, ko te tauhohenga wawe 83A:476 (6 hpi) me te tauhohenga whakaroa a Mandelup me te Taupori 22660 kei roto ko te kupu GO:0015979 'whakaahua whakaahua' me ētahi atu tukanga koiora e pā ana.
He nui rawa te whakanui i ngā kupu ontology ira tukanga koiora i tīpakohia i roto i te whakamārama o ngā ira rerekē te whakaaturanga i te wā o ngā urupare transcriptome o te lupine rau-whaiti (NLL) i werohia ki te anthrax lupine (momo Col-08 i tangohia mai i ngā mara lupine i Wierzhenice, Pōrana, i te tau 1999). Ko ngā rārangi NLL i tātarihia ko: 83A:476 (ātete, e mau ana i te ira Lanr1 homozygous), Boregine (ātete, papamuri ira kāore i te mōhiotia), Mandelup (ātete āhua, e mau ana i te ira AnMan homozygous) me te taupori 22660 (ngāwari).
Nā te mea ko te whāinga o tēnei rangahau he tautuhi i ngā ira e whai wāhi ana ki te ātete ki te anthracnose, i tātarihia ngā ira i tohua ki ngā kupu GO "GO: 0006952 Ngā urupare ārai" me "GO: 0055114 Ngā tukanga Redox" me ngā tapahi mai i te mea ko ngā toharite turanga ≥ 30 me te iti rawa kotahi te rārangi. × te pūwāhi i roto i te wā e whakakotahi ana i ngā uara hiranga tatauranga o te log2 (huringa whakapiko). Ko te maha o ngā ira i tutuki i ēnei paearu he 65 mō GO:0006952 me te 524 mō GO:0055114.
I whakaatuhia e te 83A:476 e rua ngā tihi DEG kua tāpirihia te kupu GO:0006952, ko te tuatahi i te 6 ira ia inihi (64 ira, te whakahaere ake me te heke) me te tuarua i te 24 ira ia inihi (15 ira, te whakahaere ake anake). I whakaatuhia hoki e Boregine i eke te GO:0006952 ki te tihi i te wā kotahi, engari he iti ake te DEG (11 me te 8) me te whakahohenga pai. I whakaatuhia e Mandeloop e rua ngā tihi o GO:0006952 i te 12 me te 48 HPI, e rua e mau ana i te 12 ira (ko te tuatahi me ngā ira whakahohe, me te tuarua me ngā ira pehi anake), ko te taupori 22660 i te 6 HPI (13 ira) he nui ake te mana o te pikinga o te tihi. Me mōhio ko te 96.4% o te GO:0006952 DEG i roto i ēnei tihi he rite tonu te momo urupare (ki runga, ki raro rānei), e tohu ana i te taupatupatu nui o ngā urupare parepare ahakoa ngā rerekētanga o te maha o ngā ira e whai wāhi ana. Ko te rōpū raupapatanga nui rawa atu e pā ana ki te kupu GO:0006952 e whakamārama ana i te Starvation Stress-Associated Message Protein 22 (SAM22-like), nō te akomanga 10 pathogenesis-associated protein (PR-10) protein clade me te matua protein latex. rite (MLP-like) protein (Pikitia 3). He rerekē ngā rōpū e rua i roto i te āhua o te whakaaturanga me te ahunga o te urupare. Ko ngā ira e whakamārama ana i ngā pūmua rite ki a SAM22 i whakaatu i te whakaoho pumau me te nui i ngā wā tōmua (6, 12 hpi rānei) ā, kāore i te aro i te mutunga o te whakamātautau (48 hpi), ko ngā pūmua rite ki a MLP i whakaatu i te taunga i te 6 hpi. hpi. 83A:476 me Mandelup i te 48 hp/in, tata ki ngā tohu raraunga katoa kāore i te aro. Hei tāpiri, i whai mai ngā rerekētanga i roto i ngā kōtaha whakaaturanga o ngā ira pūmua rite ki a SAM22 i te rerekētanga i roto i te ātete anthracnose, i te mea he maha ake ngā wā o ngā raina ātete e whakaoho nui ana i ēnei ira i ngā ira ngoikore ake. I whakaatu tētahi atu ira PR-10 rite ki te LlR18A/B i tētahi tauira whakapuakanga tino ōrite ki te ira pūmua rite ki te SAM22.
I tautuhia ngā wāhanga matua o te kupu tukanga koiora "GO:0006952 Defense Response" me ngā tauira whakaaturanga o ngā ira kaitono o ngā ira Lanr1 me AnMan. Ko te tauine Log2 e tohu ana i ngā uara log2 (huringa takai) i waenga i ngā tipu i werohia (Colletotrichum lupini, momo Col-08, i tangohia mai i ngā mara lupine, Wizhenica, Poland, 1999) me ngā tipu mana (i werohia ki te rūpahu) i te wā kotahi. I tātarihia ngā momo lupine rau whaiti e whai ake nei: 83A:476 (ātete, e mau ana i te ira Lanr1 homozygous), Boregine (ātete, kāore i te mōhiotia te papamuri ira), Mandelup (ātete āhua, e mau ana i te ira AnMan homozygous), me te Taupori 22660 (ngāwari).
Hei tāpiritanga, i arotakehia ngā kōtaha whakapuakanga o ngā ira kaitono RNA-seq a Lanr1 (TanjilG_05042) me AnMan (TanjilG_12861) (Pikitia 3). I whakaatu te ira TanjilG_05042 i tētahi urupare nui (whakahohenga) i te 83A:476 i te wā tuatahi anake (6 hpi), i te mea he nui te hiranga o TanjilG_12861 i roto i a Mandeloop i ngā wā e rua anake: 6 hpi (whakahaere iho) me te 24 hpi (6 hpi). Me.). whakarerekētanga) ).
Ko ngā ira tino nui rawa te whakaputa i roto i te kupu GO:0055114 "tukanga redox" ko ngā ira e whakamārama ana i ngā pūmua cytochrome P450 me te peroxidase (Pikitia 4). Mō ngā tauira i wehea mai i te 83A:476 i te 6 HPI, ko ngā uara log2 mōrahi, iti rawa rānei (huringa takai) (mō te 86.6% o ngā ira) i kitea whānuitia i waenga i ngā tipu i werohia me ngā tipu mana, e whakaatu ana i te urupare teitei o tēnei momoira ki te ira tangata i werohia. I whakaatuhia e te 83A:476 te GO: 0055114 DEG tino nui i te 6 hpi (503 ira), ko ngā toenga o ngā raina i te 48 hpi (Boregine, 31 ira; Mandelup, 85 ira; me te Taupori 22660, 78 ira)). I roto i te nuinga o ngā ira o te whānau GO:0055114, e rua ngā momo urupare ki te werohanga (whakahohenga me te aukati) i kitea. He mea whakamīharo, tae atu ki te 97.6% o ngā DEG i tautuhia mō te kupu GO: 0055114 i roto i te Mandelupe i te 48 hp. E whakaatu ana ēnei kitenga, ahakoa te iti rawa o te tauine (arā, te maha o ngā ira redox kua whakarerekētia, 85 ki te 503), ko te tauira o ngā urupare transcriptome kua whakaroahia o te mandeloup ki te anthracnose he rite ki te urupare wawe o te 83A:476. I roto i te Boregine me te Population 22660, he iti ake tēnei huihuinga i te 51.6% me te 75.6%, ia.
I whakakitea ngā tauira o te whakaaturanga o ngā wāhanga matua o te kupu o te tukanga koiora "GO:0055114 Tukanga Redox". Ko te tauine Log2 e tohu ana i ngā uara log2 (huringa takai) i waenga i ngā tipu i werohia (Colletotrichum lupini, momo Col-08, i tangohia mai i ngā mara lupine, Wizhenica, Poland, 1999) me ngā tipu mana (i werohia ki te rūpahu) i te wā kotahi. I tātarihia ngā momo lupine rau whaiti e whai ake nei: 83A:476 (ātete, e mau ana i te ira Lanr1 homozygous), Boregine (ātete, kāore i te mōhiotia te papamuri ira), Mandelup (ātete āhua, e mau ana i te ira AnMan homozygous), me te Taupori 22660 (ngāwari).
83A:476 Ko ngā urupare transcriptomic ki te werohanga ki te C. lupini (momo Col-08) i uru atu hoki ki te whakangāwari i ngā ira e pā ana ki te kupu GO:0015979 "photosynthesis" me ētahi atu tukanga koiora e pā ana (Pikitia 5). Kei roto i tēnei huinga DEG GO:0015979 he 105 ngā ira i pehia nuitia i te 6 hpi i te 83A:476. I roto i tēnei huinga iti, i whakahekehia hoki te 37 ngā ira i roto i a Mandelup i te 48 HPI me te 35 i te wā kotahi i roto i te taupori 22660, tae atu ki te 19 DEG e rite ana ki ngā momoira e rua. Kāore he DEG e pā ana ki te kupu GO: 0015979 i whakahohehia nuitia i roto i tētahi huinga (rārangi x wā).
I whakakitea ngā tauira o te whakaaturanga o ngā wāhanga matua o te tukanga koiora "GO:0015979 Photosynthesis". Ko te tauine Log2 e tohu ana i ngā uara log2 (huringa takai) i waenga i ngā tipu i werohia (Colletotrichum lupini, momo Col-08, i tangohia mai i ngā mara lupine, Wizhenica, Poland, 1999) me ngā tipu mana (i werohia ki te rūpahu) i te wā kotahi. I tātarihia ngā momo lupine rau whaiti e whai ake nei: 83A:476 (ātete, e mau ana i te ira Lanr1 homozygous), Boregine (ātete, kāore i te mōhiotia te papamuri ira), Mandelup (ātete āhua, e mau ana i te ira AnMan homozygous), me te Taupori 22660 (ngāwari).
I runga i ngā hua o te tātaritanga rerekētanga o te whakapuakanga, me te whakaaro pea he wāhi tēnei ki ngā urupare ārai ki ngā harore mate, i tīpakohia tēnei huinga ira e whitu mō te ine i ngā kōtaha whakapuakanga mā te PCR wā-tūturu (Ripanga Tāpiri S9).
I tino kitea te ira pūmua TanjilG_10657 i roto i ngā momo i rangahauhia me ngā wā katoa, ki te whakataurite ki ngā tipu mana (whakaahua) (Ripanga Tāpiri S10, S11). Hei tāpiri, i whakaatu te āhua whakapuakanga o TanjilG_10657 i tētahi ia pikinga i roto i te whakamātautau mō ngā momo katoa. I whakaatuhia e te taupori 22660 te aro nui rawa atu o TanjilG_10657 ki te werohanga me te whakahohenga 114-wā, me te taumata whakapuakanga whanaunga teitei rawa atu (4.4 ± 0.4) i te 24 HPI (Pikitia 6a). I whakaatuhia hoki e te ira pūmua PR10 LlR18A TanjilG_27015 te whakahohenga puta noa i ngā momo me ngā wā katoa, me te hiranga tatauranga i te nuinga o ngā raraunga (Pikitia 6b). Pērā i a TanjilG_10657, i kitea te taumata whakapuakanga whanaunga teitei rawa atu o TanjilG_27015 i roto i te taupori 22660 i werohangahia i te 24 HPI (19.5 ± 2.4). I tino piki ake te ira endochitinase waikawa TanjilG_04706 i ngā raina katoa me ngā wā katoa engari ko Boregine 6 hpi (Pikitia 6c). I tino whakaohokia i te wā tuatahi (6 HPI) i te 83A:476 (mā te 10.5 ngā wā) ā, i piki ake i roto i ētahi atu raina (mā te 6.6-7.5 ngā wā). I te wā o te whakamātautau, i noho tonu te whakaaturanga o TanjilG_04706 ki ngā taumata ōrite i roto i te 83A:476 me Boregine, i roto i a Mandelup me te Taupori 22660 i tino piki ake, tae atu ki ngā uara teitei (5.9 ± 1.5 me te 6.2 ± 1.5). I whakaatu te ira endoglucan-1,3-β-glucosidase-rite TanjilG_23384 i te whakahohenga teitei i ngā wā tuatahi e rua (6 me te 12 hpi) i roto i ngā raina katoa engari ko te taupori 22660 (Pikitia 6d). I kitea ngā taumata whakapuakanga whanaunga teitei rawa atu o TanjilG_23384 i te wā tuarua (12 hpi) i roto i a Mandelup (2.7 ± 0.3) me 83A:476 (1.5 ± 0.1). I te 24 HPI, he iti noa iho te whakapuakanga o TanjilG_23384 i roto i ngā rārangi katoa i rangahaua (mai i te 0.04 ± 0.009 ki te 0.44 ± 0.12).
Ngā kōtaha whakapuakanga o ngā ira kua tīpakohia (ag) i whakaaturia e te PCR tauanga. Ko ngā tau 6, 12 me te 24 e tohu ana i ngā hāora i muri i te werohanga. I whakamahia ngā ira LanDExH7 me LanTUB6 mō te whakataurite, ā, i whakamahia a LanTUB6 mō te whakataurite i waenga i ngā raupapa. Ko ngā pae hapa e tohu ana i te paerewa rerekētanga i runga i ngā tāruarua koiora e toru, ko ia o ēnei ko te toharite o ngā tāruarua hangarau e toru. Ko te hiranga tatauranga o ngā rerekētanga o ngā taumata whakaaturanga i waenga i ngā tipu i werohia (Colletotrichum lupini, momo Col-08, i riro mai i te tau 1999 mai i te mara lupin i Wierzenica, Pōrana) me ngā tipu mana (i werohia tauiratia) kua tohua i runga ake i ngā tohu raraunga (*Uara P < 0.05, **Uara P ≤ 0.01, ***Uara P ≤ 0.001). Ko te hiranga tatauranga o ngā rerekētanga o ngā taumata whakaaturanga i waenga i ngā tipu i werohia (Colletotrichum lupini, momo Col-08, i riro mai i te tau 1999 mai i te mara lupin i Wierzenica, Pōrana) me ngā tipu mana (i werohia tauiratia) kua tohua i runga ake i ngā tohu raraunga (*Uara P < 0.05, **Uara P ≤ 0.01, ***Uara P ≤ 0.001). Статистическая значимость различий в уровнях экспреспосии между инокулированными (Colletotrichum lupini, штамм Col-08, получен л. 1999 г. в Верженице, Польша) и контрольными (ложно инокулированными) растениями отмечена отмечена точками данных (*знач0наных (*знач0нание P. 0,01, ***значение P ≤ 0,001). Kei runga ake i ngā raraunga ngā rerekētanga nui ā-tatauranga i roto i ngā taumata whakaputa i waenga i ngā tipu i werohia (Colletotrichum lupini, momo Col-08, i whiwhihia i te tau 1999 mai i tētahi mara lupine i Wierzhenice, Pōrana) me ngā tipu mana (i werohia rūpahu) (*Uara P < 0.05, **Uara-P ≤ 0.01, ***Uara-P ≤ 0.001).接种（Colletotrichum lupini，Col-08株，1999年从波兰Wierzenica的羽扇豆田获得）和对照（模拟接种）植物之间表达水平差异的统计学显着性标记在数上我為0.05, **P ≤ 0.01, ***P ≤ 0.001).接种 （colletotrichum lupini ， color-08 株 ， 1999 年 波兰 波兰 wierzenica 的 羽扇 获得） 和 对照 （接秉 （接秉水平 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). Статистически значимые различия в уровнях экспрессии между инокулированными (Colletotrichum lupini, штамм Col-08, полученпли спол. Верженице, Польша, в 1999 г.) и контрольными (ложно инокулированными) растениями отмечены над точками (* данных инокулированными) P-значение ≤ 0,01, ***P-значение ≤ 0,001). Kei runga ake i ngā raraunga ngā rerekētanga nui ā-tatauranga i roto i ngā taumata whakaputa i waenga i ngā tipu i werohia (Colletotrichum lupini, momo Col-08, i tangohia mai i ngā mara lupine i Verzhenice, Pōrana, i te tau 1999) me ngā tipu mana (i werohia rūpahu) (*Uara P < 0.05 , **Uara-P ≤ 0.01, ***Uara-P ≤ 0.001).Ko ngā rārangi NLL i tātarihia ko: 83A:476 (ātete, kei a ia te ira Lanr1 homozygous), Mandelup (ātete āhua, kei a ia te ira AnMan homozygous), Boregine (ātete, kāore i te mōhiotia te papamuri ira) me te taupori 22660 (ngāwari).
I whakaatuhia e te ira kaitono a TanjilG_05042 i te wāhi Lanr1 tētahi tauira whakapuakanga rerekē rawa atu i ngā kōtaha i whiwhi mai i ngā rangahau RNA-seq (Pikitia 6e). I kitea te whakahohenga nui o tēnei ira i roto i a Mandelup me te taupori 22660 (tae atu ki te 39.7 me te 11.7 ngā wā, ia), i hua ake ai he taumata whakapuakanga teitei (tae atu ki te 1.4 ± 0.14 me te 7.2 ± 1.3, ia). I whakaatuhia hoki e te 83A:476 ētahi pikinga o te ira TanjilG_05042 (tae atu ki te 3.8-ngā wā), heoi, ko ngā taumata whakapuakanga whanaunga i tutukihia (0.044 ± 0.002) he neke atu i te 30-ngā wā iti iho i ērā i kitea i roto i a Mandelup me te taupori 22660. I tātarihia mā te qPCR, i whakaatuhia he rerekētanga nui o ngā taumata whakaaturanga i waenga i ngā momoira i roto i ngā momo i werohia ki te werohanga tauira (mana), ā, e 58 ngā wā i waenga i ngā taupori 22660 me 83A:476, tae atu ki waenga i ngā taupori 22660 me 22660. I tutuki he rerekētanga rua ngā wā i waenga i a Boregine me Mandalup.
Ko te ira kaitono i te wāhi AnMan, TanjilG_12861, i whakahohehia hei whakautu ki te werohanga i roto i te 83A:476 me Mandelup, he koretake i roto i te taupori 22660, ā, i whakahekehia i roto i te Boregine (Pikitia 6f). Ko te whakaaturanga whanaunga o te ira TanjilG_12861 te mea teitei rawa atu i roto i te 83A: 476 i werohangahia (0.14±0.01). Ko te ira pūmua ru wera akomanga I 17.4 kDa TanjilG_05080 HSP17.4 i whakaatu i ngā taumata whakaaturanga whanaunga iti iho i roto i ngā momo me ngā wā katoa i rangahauhia (Pikitia 6g). Ko te uara teitei rawa atu i kitea i te 24 HPI i roto i te taupori 22660 (0.14 ± 0.02, he pikinga waru-whakarea i roto i te urupare ki te werohanga).
I te whakataurite i ngā kōtaha whakapuakanga ira (Pikitia 7) i kitea he hononga nui i waenga i a TanjilG_10657 me ētahi atu ira e whā: TanjilG_27015 (r = 0.89), TanjilG_05080 (r = 0.85), TanjilG_05042 (r = 0.80), me TanjilG_04706 (r = 0.79). Tērā pea ka tohu ēnei hua i te whakahaere tahi o ēnei ira i te wā o ngā urupare ārai. I whakaatuhia e ngā ira TanjilG_12861 me TanjilG_23384 ngā kōtaha whakapuakanga rerekē me ngā uara tauwehenga hononga Pearson iti iho (mai i te 0.08 ki te 0.43 me te -0.19 ki te 0.28, ia) ki te whakataurite ki ētahi atu ira.
I kitea ngā hononga i waenga i ngā kōtaha whakaaturanga ira mā te whakamahi i te PCR tauanga. I tātarihia ngā momo lupine rau whaiti e whai ake nei: 83A:476 (ātete, kei a ia te ira Lanr1 homozygous), Mandelup (ātete āhua, kei a ia te ira AnMan homozygous), Boregine (ātete, kāore i te mōhiotia te papamuri ira), me te Taupori 22660 (ngāwari). E toru ngā wā i tatauhia (6, 12 me te 24 hāora i muri i te werohanga), tae atu ki ngā tipu i werohangahia (Colletotrichum lupini, momo Col-08, i riro mai i ngā mara lupine i Wierzhenice, Pōrana, i te tau 1999) me ngā tipu mana (i werohangahia ki te rūpahu). E whakaatu ana te tauine i te uara o te tauwehenga taunga Pearson.
I runga i ngā raraunga i whiwhihia i te 6 hōihopa ia inihi, i whakahaerehia te WGCNA i runga i te 9981 DEG i tautuhia mā te whakataurite i ngā tipu i werohia me ngā tipu whakahaere hei arotahi ki ngā urupare parepare wawe (Ripanga Tāpiri S12). E rua tekau mā rua ngā kōwae ira (ngā tautau) i kitea me ngā kōtaha whakaaturanga e pā ana (pai, kino rānei) i waenga i ngā momoira me ngā momo whakamātautau. I te toharite, i heke iho ngā taumata whakaaturanga ira i te raupapa 83A:476 > Mandelup > Boregine > Taupori 22660 (heoi, i ngā momo e rua, i kaha ake tēnei ia i roto i ngā tipu mana whakahaere). I te toharite, i heke iho ngā taumata whakaaturanga ira i te raupapa 83A:476 > Mandelup > Boregine > Taupori 22660 (heoi, i ngā momo e rua, i kaha ake tēnei ia i roto i ngā tipu mana whakahaere). В среднем уровни экспрессии генов снижались в порядке 83A:476 > Mandelup > Boregine > Population 22660 (i roto i te обоих вариантах, обнтакон, белет сильнее у контрольных растений). I te toharite, i heke ngā taumata whakaaturanga ira i te raupapa 83A:476 > Mandelup > Boregine > Taupori 22660 (engari, i ngā momo e rua, i kaha ake tēnei ia i roto i ngā tipu mana whakahaere).平均而言，基因表达水平按83A:476 > Mandelup > Boregine > Taupori 22660的顺序下降（然而，在两种变体中，这种趋势在对照植物中更强）。平均 而 言 ， 基因 水平 按 按 83a: 476> mandelup> boregine> taupori 22660 的 顺序 下降 （， 在 种 中 ， 在 种 中植物 中 更） . . . . . . . . . . . . . . . . . . В среднем уровни экспрессии генов снижались в ряду 83A:476 > Mandelup > Boregine > Taupori 22660 контрольных растений). I te toharite, i heke ngā taumata whakaaturanga ira i roto i te raupapa 83A:476 > Mandelup > Boregine > Taupori 22660 (heoi, i roto i ngā momo e rua, i kaha ake tēnei ia i roto i ngā tipu mana whakahaere).Nā te werohanga i hua ake ai te pikinga o te whakaaturanga ira, inā koa i roto i ngā kōwae 18, 19, 14, 6 me te 1 (i te raupapa heke o te pānga), te whakahaere kino (hei tauira, ngā kōwae 9 me te 20) me ngā pānga koretake rānei (hei tauira, ngā kōwae 11, 22, 8 me te 13). I whakaatuhia e te tātaritanga whakarei kupu GO (Ripanga Tāpiri S13) "GO: 0006952 Ngā urupare tiaki" mō te kōwae i werohia (18) me te whakahohenga mōrahi, tae atu ki ngā ira i tātarihia e te qPCR (TanjilG_04706, TanjilG_23384, TanjilG_10657 me TanjilG_27015), tae atu ki te maha o ngā kōwae whakaahua i pehia rawatia e Inoculate (9). I tautuhia te pūhui whakapūpū o te waeine 18 (Pikitia 8) hei ira TanjilG_26536 e whakamārama ana i te pūmua PR-10-rite LlR18B, ā, i tautuhia te pūhui whakapūpū o te waeine 9 hei ira TanjilG_28955 e whakamārama ana i te pūmua photosystem II PsbQ. I kitea i roto i te waeine 22 (Pikitia 9) tētahi ira ātete anthracnose Lanr1, TanjilG_05042, ā, e hono ana ki ngā kupu "GO:0044260 Cellular macromolecular metabolic processes" me "GO:0006355 Transcriptional regulation, DNA templateing" e mau ana i te TanjilG_01212 hub. Ka whakamārama te ira i te heat stress transcription factor A-4a (HSFA4a).
Tātari whatunga taumaha o te whakaaturanga ira tahi o ngā kōwae me ngā kupu tukanga koiora kua tino kitea "GO: 0006952 Ngā urupare parepare". I whakangawaritia te herenga hei whakaatu i ngā ira e whā i tātarihia e te qPCR (TanjilG_04706, TanjilG_23384, TanjilG_10657 me TanjilG_27015).
Tātari whatunga taumaha o te whakaaturanga ira tahi o tētahi kōwae me tētahi kupu tukanga koiora kua tino kitea "GO: 0006355: Te whakahaeretanga whakawhiti, te tauira DNA" me te kawe i tētahi ira ātete anthracnose Lanr1 TanjilG_05042. I whakangawaritia te herenga hei wehe i te ira TanjilG_05042 me te ira matua TanjilG_01212.
I whakaatuhia e te tirotiro ātete ki te anthracnose i kohia i Ahitereiria ko te nuinga o ngā momo i tukuna wawetia he ngoikore; kua whakaahuatia a Kalya, Coromup me Mandelup he ātete āhua, ko Wonga, Tanjil me 83A:476 he ātete tino26,27,31. he rite tonu te ira ātete, i tapaina ko Lanr1, ā, he rerekē te ira o Coromup me Mandelup, i tapaina ko AnMan10, 26, 39, i te mea i tukuna e Kalya tētahi atu ira. , Lanr2. Nā te tirotiro mō te ātete ki te anthracnose i Tiamana i kitea ai he momo ātete Bo7212 me tētahi ira kaitono ehara i te Lanr1, i tapaina ko LanrBo36.
I whakaatuhia e tā mātou rangahau he tino iti te auau (tata ki te 6%) o te ira Lanr1 i roto i te ira ira i whakamatautauria. He rite tonu tēnei kitenga ki ngā hua o te tirotiro i te ira ira o Ūropi ki te Rawhiti mā te whakamahi i ngā tohu Anseq3 me Anseq4, i whakaatu ai kei roto noa i ngā momo e rua o Belarus te ira Lanr1. E kī ana tēnei kāore anō kia whakamahia whānuitia te ira Lanr1 e ngā kaupapa whakatipu ā-rohe, he rerekē i Ahitereiria, ko tētahi o ngā ira matua mō te whakatipu ira i āwhinahia e ngā tohu. Tērā pea nā te iti o te ātete e whakaratohia ana e te ira Lanr1 i roto i ngā āhuatanga o te mara Pākehā ki te whakataurite ki te pūrongo o Ahitereiria. Hei tāpiri, kua whakaatuhia e ngā rangahau mō te anthracnose i ngā wāhi ua nui i Ahitereiria, kāore pea ngā urupare ātete e whakahaerehia ana e te ira Lanr1 e whai hua i roto i ngā āhuatanga o te rangi e pai ana ki te tipu me te whanaketanga tere o te pathogen19,42. Inaa, i roto i tēnei rangahau, i kitea anō hoki ētahi tohu o te anthracnose i roto i ngā momoira e mau ana i te ira Lanr1, e kī ana ka ngaro pea te ātete i raro i ngā āhuatanga pai mō te whanaketanga o C. lupini. Hei tāpiritanga, ka taea ngā whakamaoritanga hē mō te aroaro o ngā tohu Anseq3 me Anseq4, e tata ana ki te 1 cM mai i te wāhi Lanr1, 28,30,43.
I whakaatuhia e tā mātou rangahau ko te 83A:476, e mau ana i te ira Lanr1, i whakautu ki te werohanga C. lupini me te hōtaka anō transcriptome nui i te wā tuatahi i tātarihia (6 hpi), i roto i a Mandelup, e mau ana i te ira AnMan, i kitea ngā urupare transcriptomic i muri mai (mai i te 24 ki te 48 hp). Ko ēnei rerekētanga wā i roto i ngā urupare parepare e pā ana ki ngā rerekētanga o ngā tohu mate, e whakaatu ana i te hiranga o te mōhio wawe ki ngā pathogens mō te urupare angitu ki te ātete. Hei pangia te kiko tipu, me haere ngā spore anthrax i roto i ētahi wāhanga whanaketanga i runga i te mata o te manaaki, tae atu ki te whakatipu, te wehenga pūtau, me te hanganga o te appressorium. Ko te apiti he hanganga pangia e piri ana ki te mata o te manaaki, ā, ka āwhina i te urunga ki roto i ngā kiko manaaki. Nō reira, ko ngā spore o C. gloeosporioides i roto i te tangohanga pī i whakaatu i te wehenga tuatahi o te karihi i muri i te 75-90 meneti o te whakatipu, te hanganga o te ngongo ira i muri i te 90-120 meneti, me te pehi i muri i te 4 hāora 45. I whakaatuhia e te Mango C. gloeosporioides te neke atu i te 40% o te whakatipu conidial i muri i te 3 haora o te whakatipu me te tata ki te 20% o te hanganga o ngā kaiponu i muri i te 4 haora. Ko te ira CAP20 e pā ana ki te virulence o C. gloeosporioides i whakaatu i te mahi whakawhitiwhiti i roto i ngā conidia e hanga ana i te epiphyte i muri i te 3.5 haora o te whakatipu i roto i te ware pi o te mata o te āporo me te nui o te pūmua CAP20 i muri i te 4 haora 46 meneti. Waihoki, ko te mahi a ngā ira biosynthesis melanin i roto i te C. trifolii i whakaohohia i roto i te whakatipu 2-haora, ā, i muri mai ko te hanganga o te appressorium i muri i te 1 haora. Kua whakaatuhia e ngā rangahau o ngā kiko rau ko ngā rōpere i werohia ki te C. acutatum ka pehia tuatahi i te 8 hpi, ko ngā tōmato i werohia ki te C. coccodes ka pehia tuatahi i te 4 hpi48,49. he rite tonu ki te tauine wā o te tukanga pangia Colletotrichum spp. Ko ngā urupare tere ki te ārai mate ki te 83A:476 e kī ana ko te whai wāhi o te ātete tipu me ngā ira ārai mate e whakaohohia ana e te effector (ETI) i roto i tēnei rārangi, ko ngā urupare whakaroa a Mandelup e tautoko ana i te ariā ārai mate e whakaohohia ana e te tauira ngota ngota (MTI) 50. Ko ngā urupare wawe ki te 83A: 476 me Mandelup. Ko te taupatupatu wāhanga i waenga i ngā ira e piki ake ana, e heke iho ana rānei i roto i te urupare whakaroa e tautoko ana hoki i tēnei ariā, nā te mea e kiia ana ko te ETI he urupare MTI tere ake, e whakarei ake ana hoki, e mutu ana ki te mate pūtau kua whakaritea i te wāhi o te mate, e mōhiotia ana ko te anaphylactic shock 51,52.
Ko te nuinga o ngā ira i tapaina ki te kupu Gene Ontology GO:0006952 "Defense Response" ko ngā homologue 11 o te pūmua karere nohopuku 22 i puta mai i te ahotea (he rite ki a SAM22) me ngā momo pūmua latex matua e whitu (MLP). I whakaatu ngā pūmua rite 31, 34, 43 me te 423 i te ōrite o te raupapatanga. I whakaatu ngā ira rite SAM22 i te whakahohenga nui i roa ake te roa, e whakaatu ana i te pikinga o te ātete anthracnose (83A:476 me Boregine). Heoi, i heke iho ngā ira rite MLP i roto i ngā rārangi e mau ana i te ira ātete kaitono (83A:476/Lanr1 i te 6 hpi me Mandelup/AnMan i te 24 hpi). Me mōhio ko ngā homologue rite SAM22 katoa kua tautuhia i ahu mai i tētahi rōpū ira e horapa ana i te 105 kb, ko ngā ira rite MLP i ahu mai i ngā rohe motuhake o te ira. I kitea anō hoki te whakahohenga taunga o aua ira rite ki a SAM22 i roto i tā mātou rangahau o mua mō te ātete NLL ki te werohanga Diaporthetoxica, e kī ana kei roto rātou i ngā wāhanga whakapae o te urupare ārai. E tautokona ana hoki tēnei whakatau e ngā pūrongo mō te urupare pai o ngā ira rite ki a SAM22 ki te whara, ki te maimoatanga rānei ki te waikawa salicylic, ki ngā mea whakaoho harore, ki te hauwai waikura rānei.
Kua whakaaturia ngā ira rite ki te MLP ki te aro ki ngā ahotea abiotic me ngā ahotea biotic, tae atu ki ngā mate kitakita, huaketo me ngā mate harore pathogenic i roto i te maha o ngā momo tipu55. Ko ngā ahunga o te urupare ki ētahi taunekeneke i waenga i ngā tipu me ngā pathogens i ahu mai i te pikinga nui (arā, i te wā e pangia ana te miro e te Verticillium dahliae) ki te hekenga nui (arā, i muri i te pangia o te rākau aporo e te Alternaria spp.)56,57. Kua kitea te hekenga nui o te ira rite ki te MLP 423 i te wā e tiakina ana e te āporo te mate F. niger me te wā e pangia ana te rākau aporo Botryosphaeria berengeriana f. cn. piricola me Alternaria alternata he momo mate aporo58,59. Hei tāpiri, ko ngā kauri aporo e whakaatu nui ana i te ira rite ki te MLP 423 he iti ake te whakaaturanga o ngā ira e pā ana ki te ātete, ā, he ngawari ake ki te pangia e te mate harore59. Whai muri i te Fusarium oxysporum f, i pehia hoki te ira rite ki te MLP 423 i roto i te germplasm pīni noa ātete. cn. Mate pīni 60.
Ko ētahi atu mema o te whānau PR-10 i tautuhia i roto i tā mātou rangahau RNA-seq ko ngā ira LlR18A me LlR18B hei whakautu ki te pikinga ake o te taumata, me te ira kua piki ake (1 ira) kua heke iho rānei (3 ira) mō te pūmua whakawhiti ngako DIR1. . Hei tāpiri, e whakaatu ana a WGCNA i te ira LlR18B hei pokapū i roto i tēnei kōwae, he tino ngāwari ki te werohanga, ā, he maha ngā ira urupare tiaki kei a ia. I whakaohokia ngā ira LlR18A me LlR18B i roto i ngā rau lupine kōwhai hei whakautu ki ngā huakita mate, tae atu ki ngā kakau NLL i muri i te werohanga D. toxica, i te mea ko te ira raihi ōrite o ēnei ira, RSOsPR10, i whakaohokia teretia e te mate harore e pā ana ki te ara tohu waikawa jasmonic53,61, 62. Ka whakamāramahia e te ira DIR1 ngā pūmua kawe ngako kore-motuhake e hiahiatia ana mō te tīmatanga o te ātete whiwhi pūnaha (SAR). I te whanaketanga o ngā tauhohenga tiaki, ka haria te pūmua DIR1 mai i te pokapū o te mate mā roto i te phloem hei whakaoho i te SAR i roto i ngā okana tawhiti. He mea whakamīharo, i whakaoho nuitia te ira TanjilG_02313 DIR1 i te wā tuatahi i roto i ngā rārangi 84A:476 me te Taupori 22660, engari i angitu te whanaketanga o te ātete ki te anthracnose i roto noa i te rārangi 84A:476. Tērā pea he tohu tēnei i ētahi ngoikoretanga o te ira DIR1 i roto i te NLL, nā te mea i whakautu ngā homologues e toru e toe ana ki te werohanga i roto noa i te rārangi 83A:476 i te 6 hpi, ā, i anga iho tēnei urupare ki raro.
I roto i tā mātou rangahau, ko ngā wāhanga tino noa e pā ana ki te tukanga koiora e kiia nei ko te "GO:0055114 Tukanga Redox" ko te pūmua cytochrome P450, te peroxidase, te waikawa linoleic 9S-/13S-lipoxygenase, me te 1-aminocyclopropane-1-carboxylic acid oxidase. Hei tāpiri, e tautuhia ana e tā mātou WGCNA te homologue HSFA4a hei pokapū e kawe ana i ngā kōwae pērā i te kaitono ira ātete Lanr1 TanjilG_05042. Ko te HSFA4a he wāhanga o te whakahaere redox-dependent o te whakawhiti karihi i roto i ngā tipu.
Ko ngā pūmua Cytochrome P450 he oxidoreductases e whakakorikori ana i ngā tauhohenga hydroxylation e whakawhirinaki ana ki te NADPH me/ranei te O2 i roto i te metabolism tuatahi me te tuarua, tae atu ki te metabolism o ngā xenobiotics, tae atu ki ngā homoni, ngā waikawa ngako, ngā sterols, ngā wāhanga pakitara pūtau, ngā biopolymers, me te biosynthesis o ngā pūhui tiaki 69. I roto i tā mātou I roto i tētahi rangahau, i whakaitihia te rerekētanga o te mahi cytochrome P450 tipu mai i te -10.6 log2 (huringa whakapiko) ki te 5.7 nā te maha o ngā homologue kua whakarerekētia (37) me ngā rerekētanga o ngā tauira urupare i waenga i ngā ira motuhake, e whakaatu ana i te huringa ki runga. . Ko te whakamahi i ngā raraunga RNA-seq anake hei whakamārama i te mahi koiora e whakaarohia ana o ngā ira NLL i roto i tētahi whānau pūmua nui he tino whakapae. Heoi, he mea tika kia mōhio ko ētahi ira cytochrome P450 e hono ana ki te pikinga o te ātete ki ngā harore pathogenic, ki ngā huakita rānei, tae atu ki te takoha ki ngā tauhohenga mate pāwera69,70,71.
Ko ngā peroxidase Akomanga III he whākōkī tipu maha-mahi e uru ana ki te whānuitanga o ngā tukanga pākia i te wā e tipu ana, e whanake ana rānei ngā tipu, tae atu ki te urupare ki ngā ahotea taiao pēnei i te tote, te tauraki, te kaha o te mārama, me te whakaekenga a ngā mate72. Kei roto ngā peroxidase i te taunekeneke o ētahi momo tipu me te Anthracis, tae atu ki te Stylosanthes humilis me te C. gloeosporioides, te Lens culinaris me te C. truncatum, te Phaseolus vulgaris me te C. lindemuthianum, te Cucumis sativus me te C. lagenarium73,74,75,76. He tino tere te urupare, i ētahi wā tae noa ki te 4 HPI, i mua i te urunga o te harore ki roto i te kiko tipu73. I whakautu anō hoki te ira peroxidase ki te werohanga NLL a te D. toxica. Haunga ā rātou mahi noa ki te whakahaere i te pahūtanga waikura, te whakakore rānei i te ahotea waikura, ka taea e ngā peroxidase te whakararuraru i te tipu o ngā mate mā te hanga arai ā-tinana i runga i te whakapakari i te pakitara pūtau i te wā o te lignification, te subunit, te hononga whakawhiti rānei o ngā matū motuhake. Ka taea te kī ko tēnei mahi in silico nā te ira TanjilG_03329 e whakamārama ana i tētahi anion peroxidase hanga-lignin i piki ake i roto i tā mātou rangahau i roto i te raina ātete 83A:476 i te 6 HPI, engari kāore i roto i ētahi atu momo me ngā wā kāore i aro mai.
Ko te 9S-/13S-lipoxygenase o te waikawa linoleic te taahiraa tuatahi i roto i te ara waikura o te biosynthesis lipid78. He maha ngā mahi a ngā hua o tēnei ara i roto i te tiaki tipu, tae atu ki te whakapakari i te pakitara pūtau mā te hanganga o ngā putunga callose me te pectin, me te whakahaere i te ahotea waikura mā te whakaputa i ngā momo hāora tauhohe79,80,81,82,83. I roto i tēnei rangahau, i whakarerekētia te whakaaturanga o te waikawa linoleic 9S-/13S-lipoxygenase i roto i ngā momo katoa, engari i roto i te taupori ngoikore 22660, i puta te pikinga ake i ngā wā rerekē, i roto i ngā momo e mau ana i te ātete Lanr1 me te ira AnMan, e whakanui ana i te kanorau o te paparanga oxylipin i roto i ngā tauhohenga anthrax tiaki i waenga i ēnei momoira.
I te wā i werohia ai ki te lupine, i piki ake te kaha o te 1-aminocyclopropane-1-carboxylate oxidase (ACO) (9 ngā ira) i heke iho rānei (2 ngā ira). Haunga ngā mea e rua, i puta katoa ēnei urupare i te 6 hp. i te 83A:476. Ko te tauhohenga whākōkī i whakahaerehia e ngā pūmua ACO te taahiraa whakawhāiti tere i roto i te hanga ethylene, nō reira he tino whakahaeretia84. Ko te ethylene he homoni tipu e whai wāhi ana ki te whakahaere i te whanaketanga tipu me te urupare ki ngā āhuatanga ahotea abiotic me te biotic. Ko te whakaoho i te tuhi ACO me te whakahohenga o te ara tohu ethylene e uru ana ki te whakanui ake i te ātete o te raihi ki te harore hemibiotrophic oryzae oryzae mā te whakahaere i te hanga o ngā momo hāora tauhohe me ngā phytoalexins. He rite tonu te tukanga pangia rau i kitea i waenga i a M. oryzae me C. lupini88,89, i te taha o te pikinga nui o ngā hoa ACO i roto i te rārangi 83A:476 i kōrerotia i roto i tēnei rangahau, e neke ana i te āheinga o te tuku ātete ki te NLL anthracnose Ethylene, he taahiraa matua tohu i roto i ngā ara ngota.
I roto i tēnei rangahau, i kitea te pehitanga nui o ngā ira maha e pā ana ki te photosynthesis i te 6 hpi i roto i te 83A:476 me te 48 hpi i Mandeloop me te taupori 22660. Ko te whānuitanga me te ahunga whakamua o ēnei huringa he rite ki te taumata. I kitea te ātete ki te anthracnose i roto i tēnei whakamātautau. Nō nā tata nei, kua pūrongohia te pehitanga kaha me te wawe o ngā tuhinga e pā ana ki te photosynthesis i roto i ētahi tauira o ngā taunekeneke tipu-pathogen, tae atu ki ngā huakita pathogenic me ngā harore. Ko te tere (mai i te 2 HPI i roto i ētahi taunekeneke) me te pehitanga ā-ao o ngā ira e pā ana ki te photosynthesis hei whakautu ki te mate ka taea te whakaoho i te ārai mate tipu i runga i te whakatakotoranga o ngā momo hāora tauhohe me tā rātou taunekeneke ki te ara waikawa salicylic hei whakahaere i ngā tauhohenga mate pāwera 90,94.
Hei whakamutunga, ko ngā tikanga urupare ārai i whakaarohia mō te momo tino ātete (83A:476) ko te tere o te mōhio ki ngā mate pāwera e te ira R (tērā pea ko te TIR-NBS-LRR TanjilG_05042) me te tohu waikawa salicylic me te ethylene i tukuna e te urupare mate pāwera, ā, muri iho ko te whakatūnga o te SAR tawhiti. Ka tautokona te mahi e te pūmua DIR-1. Me mōhio he poto rawa te wā biotrophic mō te mate C. lupini (tata ki te 2 rā), ā, muri iho ko te tipu necrotic95. Ko te whakawhiti i waenga i ēnei wāhanga ka hono pea ki te necrosis me te whakaaturanga o ngā pūmua i puta mai i te ethylene e mahi ana hei whakaoho i ngā tauhohenga hypersensitivity i roto i ngā tipu manaaki. Nō reira, he tino whaiti te matapihi wā mō te hopu angitu o C. lupini i te wāhanga biotrophic. Ko te hōtaka anō o ngā ira e hono ana ki te redox me te photosynthesis i kitea i roto i te 83A:476 i te 6 hpi e rite ana ki te haere whakamua o te hyphae harore, ā, e tohu ana i te whanaketanga o te urupare tiaki angitu i te wāhanga biotrophic. Tērā pea kua roa rawa ngā urupare transcriptomic a Mandelup me te taupori 22660 ki te hopu i te harore i mua i te huri ki te tipu necrotic, heoi, tera pea he whai hua ake a Mandelup i te taupori 22660 nā te mea ka whakatairangahia e te whakahaere tere o te pūmua PR-10 te ātete whakapae.
Ko te ETI, e ārahihia ana e te ira R matua, te āhua nei he tikanga noa mō te ātete pīni ki te anthracnose. Nō reira, i roto i te tauira pīni Medicago truncatula, ko te ātete ki te anthracnose ka tukuna e te ira RCT1, he mema nō te akomanga ira R tipu TIR-NBS-LRR97. Ka tukuna hoki e tēnei ira te ātete whānui ki te anthracnose i roto i te alfalfa ina whakawhitia ki ngā tipu ngoikore. I roto i te pīni noa (P. vulgaris), neke atu i te rua tekau mā rua ngā ira ātete ki te anthracnose kua tautuhia tae noa ki tēnei wā. Ko ētahi o ēnei ira ka kitea i ngā rohe kāore he ira R matua, engari he maha atu kei ngā taha o ngā chromosomes e mau ana i te tautau ira NBS-LRR, tae atu ki te TIR-NBS-LRRs99. I whakaūhia hoki e te rangahau SSR whānui-ira te hononga o te ira NBS-LRR me te ātete ki te anthracnose i roto i te pīni noa. I kitea anō hoki te ira R matua i roto i te rohe ira e mau ana i te wāhi ātete nui ki te anthracnose i roto i te lupine mā 101.
E whakaatu ana tā mātou mahi he pai te tiaki i te lupine rau-whaiti mai i te anthracnose i puta mai i te harore mate pāwera Collelotrichum lupini mā te whakamahi i te raupapatanga tere, i whakaatu mātou i ngā momo whakaaturanga rerekē o ngā ira ātete anthracnose i roto i ngā tipu NLL i whakahaerehia e ngā ira ātete Lanr1 me AnMan. Ko te parenga angitu ko te hoahoa āta i ngā ira mō ngā pūmua e uru ana ki te redox, te photosynthesis, me te pathogenesis i roto i ngā hāora o te whakapā tuatahi o te tipu ki te pathogen. Ko ngā tauhohenga tiaki rite tonu, engari i roa te wā, he iti ake te whai huatanga ki te tiaki i ngā tipu mai i ngā mate. Ko te ātete anthrax i whakahaerehia e te ira Lanr1 he rite ki te urupare tere noa o te ira R (te ārai mate i puta mai i te effector), ko te ira AnMan he mea pea ka whakarato i te urupare whakapae (te ārai mate i puta mai i te tauira ngota ngota e hono ana ki te microbe), e whakarato ana i te taumata pumau taurite.
Ko ngā rārangi NLL e 215 i whakamahia hei tirotiro i ngā tohu anthracnose he 74 ngā momo, 60 ngā rārangi i whiwhi mā te whakawhiti, te whakaputa uri rānei, 5 ngā momo ira whakarerekē, me te 76 ngā ira tūturu, taketake rānei. I ahu mai ngā rārangi i ngā whenua 17, ko te nuinga i Pōrana (58), Pāniora (47), Tiamana (27), Ahitereiria (26), Rūhia (19), Peraruhia (7), Itari (5) me ētahi atu rārangi. mai i ngā whenua 10. Kei roto hoki i te huinga ngā rārangi ātete tohutoro: 83A:476, Tanjil, Wonga e mau ana i te ira Lanr1, me Mandelup e mau ana i te ira AnMan. I riro mai ngā rārangi mai i te Pūtake Raraunga Rauemi Ira Lupine Pākehā e puritia ana e Poznań Plant Breeding Ltd., Wiatrowo, Pōrana (Ripanga Tāpiri S1).
I whakatipuhia ngā tipu i raro i ngā āhuatanga whakahaere (16 hāora te roa o te wā whakaahua, 25°C te pāmahana i te awatea, 18°C ​​i te pō). E rua ngā tauira koiora i tātarihia. I wehea te DNA mai i ngā rau e toru wiki te pakeke mā te whakamahi i te DNeasy Plant Mini Kit (Qiagen, Hilden, Tiamana) e ai ki te tikanga. I aromatawaihia te kounga me te kukū o te DNA wehea mā ngā tikanga ine-whakaahua (NanoDrop 2000; Thermo Fisher Scientific, Waltham, MA, USA). I ​​tātarihia te tohu AnManM1 e tohu ana i te ira ātete anthracnose AnMan (i ahu mai i cv. Mandelup) me ngā tohu Anseq3 me Anseq4 e taha ana i te ira Lanr1 (i ahu mai i cv. Tanjil) 11,26,28. I tohua ngā homozygotes mō te ira ātete hei "1", ngā ngāwari - hei "0", me ngā heterozygotes - hei 0.5.
I runga i ngā hua o te tirotiro mō ngā tohu AnManM1, AnSeq3 me AnSeq4 me te wātea o ngā purapura mō ngā whakamātautau whai muri whakamutunga, i tīpakohia e 50 ngā momo NLL mō te tohu ātete ki te anthracnose. I mahia te tātari i roto i ngā tāruarua i roto i tētahi whare kati kōtuhi e whakahaerehia ana e te rorohiko me te wā whakaahua 14 hāora me te awhe pāmahana o te 22°C i te awatea me te 19°C i te pō. Ka waruhia ngā purapura (ka tapahia te kiri purapura i te taha ke o te kukune ki te mata koi) i mua i te rui hei ārai i te moe o te purapura nā te mea he pakeke rawa te kiri purapura, ā, kia rite ai te whakatipu. I whakatipuhia ngā tipu ki roto i ngā kohua (11 × 11 × 21 cm) me te oneone parakore (TS-1 REC 085 Medium Basic, Klasmann-Deilmann Polska, Warsaw, Pōrana). I whakahaerehia te werohanga ki te momo Colletotrichum lupini Col-08, i whakatipuria i te tau 1999 mai i ngā kakau o ngā tipu lupine rau-whaiti i whakatipuria i roto i tētahi mara i Verzhenitsa, Pōrana Nui (52° 27′ 42″ N 17° 04′ 05″ E). Tikina he wāhi. I whakatipuria ngā momo i roto i te reo SNA i te 20° C i raro i te mārama pango mō ngā rā 21 hei whakaoho i te whakapūngao. E whā wiki i muri i te rui, i te taenga o ngā tipu ki te wāhanga rau 4-6, i whakahaerehia te werohanga mā te rehu ki te whakatārewatanga o ngā conidia i te kukū o te 0.5 x 106 conidia ia ml. I muri i te werohanga, i puritia ngā tipu ki te pōuri mō ngā hāora 24 i te haumākū o te 98% me te pāmahana o te 25°C hei whakahaere i te pihinga o ngā conidia me te tukanga pangia. Kātahi ka whakatipuria ngā tipu i raro i te wā whakaahua 14-hāora i te 22°C i te awatea/19°C i te pō me te haumākū 70%. I hangaia te kaute mate i ngā rā 22 i muri i te werohanga, ā, i ahu mai i te 0 (ātete) ki te 9 (tino pāngia) i runga i te aroaro, te ngaro rānei o ngā whara necrotic i runga i ngā kakau me ngā rau. Hei tāpiri, i muri i te kaute, i inehia te taumaha o ngā tipu. I tatauhia ngā whanaungatanga i waenga i ngā momoira tohu me ngā momoira mate hei hononga ira rua-raupapa (te korenga o ngā tohu heterozygous i roto i te huinga rārangi hei tātari i te momoira ātete anthracnose).