Characterization of RNA silencing components in the plant pathogenic fungus Fusarium graminearum.
Characterization of RNA silencing components in the plant pathogenic fungus Fusarium graminearum.
复制标题
植物病原真菌禾谷镰刀菌 RNA 沉默成分的表征
DOI：
10.1038/srep12500
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发表时间：
2015-07-27
影响因子：
4.6
通讯作者：
Ma Z
中科院分区：
文献类型：
--
作者：
Chen Y;Gao Q;Huang M;Liu Y;Liu Z;Liu X;Ma Z
RNA干扰（RNA interference，RNAi）在多种真核生物的基因调控中发挥着重要作用。然而，RNAi在植物病原真菌中的作用在很大程度上仍不清楚。在这项研究中，我们探讨了RNAi途径的核心组件在小麦赤霉病的主要致病因子禾谷镰刀菌中的作用。我们的研究结果表明，发夹RNA（hpRNA）可以有效地沉默目的基因的表达水平，argonaute蛋白FgAgo 1和Dicer蛋白FgDicer 2在这一沉默过程中起着重要作用。RNAi机制不参与F.在试验条件下，本研究首次应用高通量测序技术对F.结果发现，在野生型和FgDicer 2中鉴定了总共49种微样RNA（milRNA）候选物，其中24种是FgDicer 2依赖性的。Fg-milRNA-4负调节其靶基因的表达。以上结果表明，hpRNA诱导的基因沉默是一种有价值的研究基因功能的遗传工具。禾谷早熟禾FgAgo 1和FgDicer 2蛋白在hpRNA介导的基因沉默过程中起关键作用。此外，FgDicer 2还参与了该真菌sRNA的转录和milRNA的产生。
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.