本文關鍵詞： 大豆花葉病毒 大豆 microRNA 抗病基因 RNAi途徑 微陣列分析　出處：《內(nèi)蒙古大學》2016年博士論文　論文類型：學位論文

【摘要】：植物通過不同層次的防衛(wèi)反應來抵御外源病毒的侵染。RNA干擾(RNA interference, RNAi)是植物針對病毒侵染的一個重要的應答反應,AGO (Argonaute)和Dicer/DCL (Dicer-like)蛋白是RNAi途徑中的關鍵成分。效應蛋白誘導的免疫反應(Effector triggered immunity, ETI)一般指的是抗病基因(Resistance gene, R)介導的抗病毒機制。迄今為止發(fā)現(xiàn)的R蛋白多數(shù)屬于NBS-LRR家族蛋白。MicroRNA (miRNA)是一類由生物體內(nèi)源產(chǎn)生的、具有重要調(diào)節(jié)作用的非編碼小RNA分子。近年來發(fā)現(xiàn)miRNA在植物的抗病反應中也發(fā)揮著很重要的作用。大豆花葉病毒(Soybean mosaic virus, SMV)是一種世界性大豆病害,嚴重影響著大豆的經(jīng)濟和商品價值。有研究表明miRNA在大豆種粒斑駁形成過程發(fā)揮著主要作用,同時發(fā)現(xiàn)SMV的侵染影響大豆內(nèi)源小干擾RNA (small interference RNA, siRNA)的積累水平。但是,目前沒有一個系統(tǒng)的研究SMV侵染與大豆miRNA表達調(diào)控之間關系的報道。本論文中我們利用microarray技術在敏感大豆品系中篩選到了45個SMV響應的miRNAs。在上調(diào)的miRNAs中miR168a、miR403a、miR162b和miR1515a的預測靶標分別為AGO1、AGO2、DCL1和DCL2; miR1507a、miR1507c和miR482a的預測靶標為NBS-LRR家族的10個抗病基因。我們通過雙熒光素酶報告系統(tǒng)與RLM-RACE實驗證實了上述7個miRNAs對其10個靶標基因表達的調(diào)控作用。AGO1、DCL1、DCL2和5個NBS-LRR類抗病基因的表達受SMV侵染的抑制,同時相應miRNAs的豐度上調(diào),并且它們之間的表達在SMV侵染后不同時間點均表現(xiàn)出相反的表達模式。我們的結果表明SMV會通過上調(diào)一些miRNAs來抑制RNAi途徑相關基因及NBS-LRR家族抗病基因的表達,從而導致敏感大豆品系的感病。
[Abstract]:Plant defense response to foreign virus infection by different levels. RNA interference RNA interference (RNAi) is an important response response to virus infection in plants ago and Dicer/DCL Dicer-like proteins are the key components of the RNAi pathway. The immune response induced by Effector triggered (ETI) is generally referred to as the antiviral mechanism mediated by resistance gene resistance (RIA). Most of the R proteins found so far belong to NBS-LRR family proteins. MicroRNAs are produced from biological sources. Noncoding small RNA molecules with important regulatory functions. In recent years, miRNA has been found to play an important role in plant disease response. Soybean mosaic virus (Soy bean mosaic virus) is a worldwide soybean disease. Some studies have shown that miRNA plays a major role in the formation of soybean seed mottles, and it is also found that the infection of SMV affects the accumulation level of small interference RNAs (siRNAs) in soybean. There is no systematic study on the relationship between SMV infection and the regulation of miRNA expression in soybean. In this paper, we used microarray technique to screen 45 SMV responsive miRNAs in sensitive soybean strains. The predicted targets were AGO1, AGO2, DCL1 and DCL2, respectively, and the predicted targets of miR1507a- miR1507c and miR482a were 10 resistant genes of the NBS-LRR family. We confirmed the regulatory effect of these 7 miRNAs on the expression of their 10 target genes by using double luciferase reporting system and RLM-RACE experiments. The expression of DCL2 and 5 NBS-LRR class resistance genes were inhibited by SMV infection. At the same time, the abundance of miRNAs was upregulated. Our results showed that SMV could inhibit the expression of RNAi pathway related genes and NBS-LRR family resistance genes by up-regulating some miRNAs. Thus, sensitive soybean strains are susceptible to the disease.
【學位授予單位】：內(nèi)蒙古大學
【學位級別】：博士
【學位授予年份】：2016
【分類號】：S435.651

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本文編號：1526117