Travella Silvia, Keller Beat
Institute of Plant Biology, University of Zürich, Zürich, Switzerland.
Methods Mol Biol. 2009;478:185-99. doi: 10.1007/978-1-59745-379-0_12.
Down-regulation of endogenous genes via post-transcriptional gene silencing (PTGS) is a key to the characterization of gene function in plants. Many RNA-based silencing mechanisms such as post-transcriptional gene silencing, co-suppression, quelling, and RNA interference (RNAi) have been discovered among species of different kingdoms (plants, fungi, and animals). One of the most interesting discover ies was RNAi, a sequence-specific gene-silencing mechanism initiated by the introduction of double-stranded RNA (dsRNA), homologous in sequence to the silenced gene, which triggers degradation of mRNA. Infection of plants with modified viruses can also induce RNA silencing and is referred to as virus-induced gene silencing (VIGS). In contrast to insertional mutagenesis, these emerging new reverse genetic approaches represent a powerful tool for exploring gene function and for manipulating gene expression experimentally in cereal species such as barley and wheat. We examined how RNAi and VIGS have been used to assess gene function in barley and wheat, including molecular mechanisms involved in the process and available methodological elements, such as vectors, inoculation procedures, and analysis of silenced phenotypes.
通过转录后基因沉默（PTGS）下调内源基因是植物基因功能表征的关键。在不同界（植物、真菌和动物）的物种中发现了许多基于RNA的沉默机制，如转录后基因沉默、共抑制、基因压制和RNA干扰（RNAi）。最有趣的发现之一是RNAi，这是一种序列特异性基因沉默机制，由引入与沉默基因序列同源的双链RNA（dsRNA）引发，进而触发mRNA的降解。用修饰病毒感染植物也可诱导RNA沉默，这被称为病毒诱导的基因沉默（VIGS）。与插入诱变不同，这些新兴的新反向遗传学方法是探索基因功能以及在大麦和小麦等谷物物种中通过实验操纵基因表达的有力工具。我们研究了RNAi和VIGS如何用于评估大麦和小麦中的基因功能，包括该过程涉及的分子机制以及可用的方法要素，如载体、接种程序和沉默表型分析。
