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Author:
Date:
2016-01-20
[Abstract] Virus-based microRNA silencing (VbMS) is a viable and prompt method to screen and characterize the function of microRNAs (miRNAs) in plants. The Tobacco rattle virus (TRV)-based VbMS method was originally developed by the Yule Liu's group (Sha et al., 2014) using miRNA target mimic (TM) methodology. Here, we describe the TRV-based VbMS method for silencing endogenous miRNA in Nicotiana benthamiana and tomato via Agrobacterium infiltrations. For each assay, Agrobacterium cultures containing pTRV1 and specific pTRV2e derivative harboring TM fragments are mixed and infiltrated into plant tissues. Generally within 3 weeks, the target miRNAs gene will be silenced and the newly developed tissues will exhibit corresponding phenotypes.
[摘要] 基于病毒的微RNA沉默（VbMS）是筛选和表征微RNA（miRNA）在植物中的功能的可行和迅速的方法。 基于烟草响铃病毒（TRV）的VbMS方法最初是由Yule Liu的小组（Sha等人，2014）使用miRNA靶模拟物TM方法开发的。 在这里，我们描述基于TRV的VbMS方法，用于通过土壤杆菌浸润物在本氏烟草和番茄中沉默内源性miRNA。 对于每个测定，将含有pTRV1的土壤杆菌培养物和含有TM片段的特异性pTRV2e衍生物混合并渗入植物组织中。 通常在3周内，靶miRNA基因将被沉默，并且新开发的组织将显示相应的表型。
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