| Virus-based MicroRNA Silencing
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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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| VIGS Assays
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Author:
Date:
2014-03-05
[Abstract] Virus-induced gene silencing (VIGS) is a powerful method to study gene function in plants. Tobacco rattle virus (TRV)-based VIGS vector is the most efficient VIGS vector so far. This method was originally developed by the Dinesh-Kumar's group (Liu et al., 2002) . Here, we describe a rapid and high efficient TRV-based VIGS method for knocking down genes in Nicotiana benthamiana. For TRV-based VIGS, Agrobacterium culture containing pTRV1 and Agrobacterium culture containing pTRV2 with plant target gene fragment are mixed and infiltrated into the lower leaves of plant. After 2-3 weeks post infiltration, plant target gene will be silenced.
[摘要] 病毒诱导基因沉默（VIGS）是研究植物中基因功能的有力方法。 迄今为止，基于烟草瘟病毒（TRV）的VIGS载体是最有效的VIGS载体。 该方法最初由Dinesh-Kumar的小组开发（Liu等人，2002）。 在这里，我们描述了一种快速和高效的基于TRV的VIGS方法，用于在本氏烟草中敲除基因。 对于基于TRV的VIGS，将含有具有植物靶基因片段的pTRV2的pTRV1和农杆菌培养物的农杆菌培养物混合并渗入植物的下叶。 在浸润后2-3周后，植物靶基因将被沉默。
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