真菌蛋白激发子MoHrip2诱导水稻抗病性的分子机制
发布时间：2019-06-15 09:51
【摘要】：Mo Hrip2是从稻瘟病菌中分离得到的一种蛋白激发子,能够增强水稻对抗稻瘟病的特性。为了阐明MoHrip2诱导水稻抗病性的分子机制,本研究将MoHrip2喷施处理水稻幼苗叶片后,检测处理不同时间后相关抗性转录因子(transcription factors,TFs)、水杨酸(SA)和茉莉酸(JA)信号传导途径的标志基因、病程相关基因(pathogenesis-related(PR)的表达水平,并测定了水稻叶片内源SA和JA的含量。利用双向电泳研究了Mo Hrip2处理24 h后水稻叶片的差异蛋白表达谱。qPCR结果表明,选定的TFs均在处理后2 h诱导表达,处理后4h表达量最高。SA和JA信号传导途径相关基因(OsNPR1/Os NH1,Os PAL1和Os LOX2)也在处理后2 h上调表达,而Os EDS1和Os AOS2下调表达。PR基因(OsPR1a,OsPR2,OsPR3,OsPR4,OsPR5和OsPR10)也在激发子处理(2-48 h)后均上调表达;葡聚糖基因Os PR2和几丁质酶基因OsPR3和OsPR4在处理2 4 h后表达量达到最高,其它的PR基因OsPR1a,OsPR5和OsPR10处理24 48 h后表达量最高。此外,激发子诱导的水稻叶片中内源SA和JA含量比未处理的明显提高,更进一步证实了SA和JA传导途径相关基因上调表达的结果。双向电泳结果显示,与对照缓冲液处理组的结果相比,Mo Hrip2处理组中诱导产生了10个新蛋白,4个上调蛋白和3个下调蛋白。这17个差异表达蛋白经MS/MS检测分析,可根据它们的假定注释功能分成六类:防御相关的转录因子,信号传导相关蛋白,活性氧组分(ROS),细胞程序性死亡(PCD),防御相关蛋白以及光合作用和能量相关蛋白。qPCR结果进一步验证了双向电泳获得的差异蛋白。采用蛋白互作试剂盒(Thermo Scientific)捕获到了4个MoHrip2的候选结合蛋白,并克隆了这些蛋白的编码基因,从体内和体外验证蛋白互作。生物信息学分析表明,Mo Hrip2可能直接或间接与过氧化物酶(ROS-相关蛋白)相互作用,参与诱导的抗病信号传导。总之,Mo Hrip2通过压力相关途径激发水稻幼苗早期防御反应,诱导PR基因上调表达,激活SA和JA信号传导途径,最终诱导水稻提高抗病性。本研究结果为水稻遗传育种和科学阐述激发子诱导水稻抗病分子机制提供了理论基础。
[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，
本文编号：2500132
[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，
本文编号：2500132
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