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植物病原真菌禾谷镰刀菌中RNA沉默组分的鉴定

Characterization of RNA silencing components in the plant pathogenic fungus Fusarium graminearum.

作者信息

Chen Yun, Gao Qixun, Huang Mengmeng, Liu Ye, Liu Zunyong, Liu Xin, Ma Zhonghua

机构信息

Institute of Biotechnology, Zhejiang University, Hangzhou, 310058, China.

出版信息

Sci Rep. 2015 Jul 27;5:12500. doi: 10.1038/srep12500.

Abstract

The RNA interference (RNAi) plays a critical role in gene regulation in a variety of eukaryotic organisms. However, the role of RNAi remains largely unclear in plant pathogenic fungi. In this study, we explored the roles of core components of the RNAi pathway in Fusarium graminearum, the major causal agent of wheat head blight. Our results demonstrated that the hairpin RNA (hpRNA) can efficiently silence the expression level of target gene, and the argonaute protein FgAgo1 and dicer protein FgDicer2 are important in this silencing process. RNAi machinery was not involved in growth, abiotic stress and pathogenesis in F. graminearum under tested conditions. We firstly applied high-throughput sequencing technology to elucidate small RNA (17-40 nucleotides) (sRNA) transcriptome in F. graminearum, and found that a total of forty-nine micro-like-RNA (milRNA) candidates were identified in the wild-type and ∆FgDICER2, and twenty-four of them were FgDicer2-dependent. Fg-milRNA-4 negatively regulated expression of its target gene. Taken together, our results indicated that the hpRNA-induced gene silencing was a valuable genetic tool for exploring gene function in F. graminearum. FgAgo1 and FgDicer2 proteins played a critical role in the hpRNA mediated gene silencing process. In addition, FgDicer2 was involved in sRNA transcription and milRNA generation in this fungus.

摘要

RNA干扰(RNAi)在多种真核生物的基因调控中发挥着关键作用。然而,RNAi在植物病原真菌中的作用仍 largely不清楚。在本研究中,我们探究了RNAi途径的核心组分在小麦赤霉病的主要病原菌禾谷镰刀菌中的作用。我们的结果表明,发夹RNA(hpRNA)能够有效地沉默靶基因的表达水平,并且AGO蛋白FgAgo1和Dicer蛋白FgDicer2在该沉默过程中很重要。在测试条件下,RNAi机制不参与禾谷镰刀菌的生长、非生物胁迫和致病过程。我们首先应用高通量测序技术来阐明禾谷镰刀菌中的小RNA(17 - 40个核苷酸)(sRNA)转录组,并发现野生型和∆FgDICER2中共鉴定出49个类微小RNA(milRNA)候选物,其中24个依赖于FgDicer2。Fg - milRNA - 4负调控其靶基因的表达。综上所述,我们的结果表明hpRNA诱导的基因沉默是探索禾谷镰刀菌基因功能的一种有价值的遗传工具。FgAgo1和FgDicer2蛋白在hpRNA介导的基因沉默过程中发挥着关键作用。此外,FgDicer2参与了该真菌中的sRNA转录和milRNA产生。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1b51/4515635/93cff6fbbaf4/srep12500-f1.jpg

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