【摘要】：Mo Hrip2是從稻瘟病菌中分離得到的一種蛋白激發(fā)子,能夠增強水稻對抗稻瘟病的特性。為了闡明MoHrip2誘導(dǎo)水稻抗病性的分子機制,本研究將MoHrip2噴施處理水稻幼苗葉片后,檢測處理不同時間后相關(guān)抗性轉(zhuǎn)錄因子(transcription factors,TFs)、水楊酸(SA)和茉莉酸(JA)信號傳導(dǎo)途徑的標(biāo)志基因、病程相關(guān)基因(pathogenesis-related(PR)的表達水平,并測定了水稻葉片內(nèi)源SA和JA的含量。利用雙向電泳研究了Mo Hrip2處理24 h后水稻葉片的差異蛋白表達譜。qPCR結(jié)果表明,選定的TFs均在處理后2 h誘導(dǎo)表達,處理后4h表達量最高。SA和JA信號傳導(dǎo)途徑相關(guān)基因(OsNPR1/Os NH1,Os PAL1和Os LOX2)也在處理后2 h上調(diào)表達,而Os EDS1和Os AOS2下調(diào)表達。PR基因(OsPR1a,OsPR2,OsPR3,OsPR4,OsPR5和OsPR10)也在激發(fā)子處理(2-48 h)后均上調(diào)表達;葡聚糖基因Os PR2和幾丁質(zhì)酶基因OsPR3和OsPR4在處理2 4 h后表達量達到最高,其它的PR基因OsPR1a,OsPR5和OsPR10處理24 48 h后表達量最高。此外,激發(fā)子誘導(dǎo)的水稻葉片中內(nèi)源SA和JA含量比未處理的明顯提高,更進一步證實了SA和JA傳導(dǎo)途徑相關(guān)基因上調(diào)表達的結(jié)果。雙向電泳結(jié)果顯示,與對照緩沖液處理組的結(jié)果相比,Mo Hrip2處理組中誘導(dǎo)產(chǎn)生了10個新蛋白,4個上調(diào)蛋白和3個下調(diào)蛋白。這17個差異表達蛋白經(jīng)MS/MS檢測分析,可根據(jù)它們的假定注釋功能分成六類:防御相關(guān)的轉(zhuǎn)錄因子,信號傳導(dǎo)相關(guān)蛋白,活性氧組分(ROS),細(xì)胞程序性死亡(PCD),防御相關(guān)蛋白以及光合作用和能量相關(guān)蛋白。qPCR結(jié)果進一步驗證了雙向電泳獲得的差異蛋白。采用蛋白互作試劑盒(Thermo Scientific)捕獲到了4個MoHrip2的候選結(jié)合蛋白,并克隆了這些蛋白的編碼基因,從體內(nèi)和體外驗證蛋白互作。生物信息學(xué)分析表明,Mo Hrip2可能直接或間接與過氧化物酶(ROS-相關(guān)蛋白)相互作用,參與誘導(dǎo)的抗病信號傳導(dǎo)�？傊�,Mo Hrip2通過壓力相關(guān)途徑激發(fā)水稻幼苗早期防御反應(yīng),誘導(dǎo)PR基因上調(diào)表達,激活SA和JA信號傳導(dǎo)途徑,最終誘導(dǎo)水稻提高抗病性。本研究結(jié)果為水稻遺傳育種和科學(xué)闡述激發(fā)子誘導(dǎo)水稻抗病分子機制提供了理論基礎(chǔ)。
[Abstract]:Mo Hrip2 is a protein elicitor isolated from Magnaporthe grisea, which can enhance the resistance of rice to rice blast. In order to elucidate the molecular mechanism of rice disease resistance induced by MoHrip2, the related resistance transcription factors (transcription factors,TFs), marker genes of salicylic acid (SA) and jasmonic acid (JA) signal transduction pathway and the expression level of disease course related genes (pathogenesis-related (PR) in rice seedling leaves were detected after MoHrip2 spraying for different time, and the contents of endogenous SA and JA in rice leaves were determined. The differential protein expression profiles of rice leaves treated with Mo Hrip2 for 24 h were studied by two-dimensional electrophoresis. The results of qPCR showed that the selected TFs was induced to express at 2 h after treatment, and the highest expression was observed at 4 h after treatment. Sa and JA signal transduction pathway related genes (OsNPR1/Os NH1,Os PAL1 and Os LOX2) were also up-regulated at 2 h after treatment, while Os EDS1 and Os AOS2 down-regulated the expression of PR gene (OsPR1a,OsPR2,OsPR3,OsPR4,). OsPR5 and OsPR10 were also upregulated after elicitor treatment (2 鈮，

本文編號：2500132