State Key Laboratory for Agro-Biotechnology and Department of Plant Pathology, China Agricultural University, Beijing, China.
Methods Mol Biol. 2022;2400:149-161. doi: 10.1007/978-1-0716-1835-6_15.
As an efficient tool for functional genomics, VIGS (virus-induced gene silencing) has been widely used in reverse and forward genetics to identify genes involved in various biology processes in many plant species. Up to now, at least 50 VIGS vectors based on RNA viruses, DNA viruses or satellites have been developed for either dicots or monocots or both. Silencing specific genes using VIGS vector involves five major steps including, first, choosing an appropriate VIGS vector for the plant; second, selecting a fragment of targeted host gene; third, cloning the fragment into viral VIGS vector; forth, inoculating and infecting the appropriate plant; and fifth, quantifying silencing effects including recording silencing phenotypes and determining silencing efficiency of the target gene. In this chapter, we introduce these steps for VIGS assay in dicots and monocots, by taking a cucumber mosaic virus-based VIGS vector for Nicotiana benthamiana and maize plants as an example. Moreover, we list available VIGS vectors for monocots.
作为功能基因组学的有效工具,VIGS(病毒诱导的基因沉默)已被广泛应用于反向和正向遗传学,以鉴定参与多种植物生物学过程的基因。迄今为止,已经开发了至少 50 种基于 RNA 病毒、DNA 病毒或卫星的 VIGS 载体,用于双子叶植物或单子叶植物或两者。使用 VIGS 载体沉默特定基因涉及五个主要步骤,包括:首先,为植物选择合适的 VIGS 载体;其次,选择目标宿主基因的片段;第三,将片段克隆到病毒 VIGS 载体中;第四,接种和感染适当的植物;第五,量化沉默效果,包括记录沉默表型和确定目标基因的沉默效率。在本章中,我们以基于黄瓜花叶病毒的 VIGS 载体为例,介绍双子叶植物和单子叶植物 VIGS 测定的这些步骤。此外,我们列出了单子叶植物可用的 VIGS 载体。