Sha Aihua, Zhao Jinping, Yin Kangquan, Tang Yang, Wang Yan, Wei Xiang, Hong Yiguo, Liu Yule
MOE Key Laboratory of Bioinformatics, Center for Plant Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China.
Plant Physiol. 2014 Jan;164(1):36-47. doi: 10.1104/pp.113.231100. Epub 2013 Dec 2.
MicroRNAs (miRNAs) play pivotal roles in various biological processes across kingdoms. Many plant miRNAs have been experimentally identified or predicted by bioinformatics mining of small RNA databases. However, the functions of these miRNAs remain largely unknown due to the lack of effective genetic tools. Here, we report a virus-based microRNA silencing (VbMS) system that can be used for functional analysis of plant miRNAs. VbMS is performed through tobacco rattle virus-based expression of miRNA target mimics to silence endogenous miRNAs. VbMS of either miR172 or miR165/166 caused developmental defects in Nicotiana benthamiana. VbMS of miR319 reduced the complexity of tomato (Solanum lycopersicum) compound leaves. These results demonstrate that tobacco rattle virus-based VbMS is a powerful tool to silence endogenous miRNAs and to dissect their functions in different plant species.
微小RNA(miRNA)在各生物界的多种生物学过程中发挥着关键作用。许多植物miRNA已通过对小RNA数据库进行生物信息学挖掘而得到实验鉴定或预测。然而,由于缺乏有效的遗传工具,这些miRNA的功能在很大程度上仍不为人知。在此,我们报道了一种基于病毒的微小RNA沉默(VbMS)系统,该系统可用于植物miRNA的功能分析。VbMS通过基于烟草脆裂病毒表达miRNA靶标模拟物来沉默内源性miRNA。对miR172或miR165/166进行VbMS会导致本氏烟草出现发育缺陷。对miR319进行VbMS降低了番茄(Solanum lycopersicum)复叶的复杂度。这些结果表明,基于烟草脆裂病毒的VbMS是一种强大的工具,可用于沉默内源性miRNA并剖析其在不同植物物种中的功能。