Scofield Steven R, Brandt Amanda S
Department of Agronomy, Purdue University, West Lafayette, IN, USA.
Methods Mol Biol. 2012;894:93-112. doi: 10.1007/978-1-61779-882-5_7.
Virus-induced gene silencing (VIGS) is a useful functional genomics tool for rapidly creating plant gene knockout phenotypes that can be used to infer gene function. Until recently, VIGS has only been possible in dicotyledonous plants. However, the development of cloning vectors based on Barley stripe mosaic virus (BSMV) has now made VIGS possible in barley and wheat. VIGS has particular advantages for functional genomics in wheat, where the organism's hexaploidy and recalcitrance to transformation have greatly hindered strategies for the functional identification of genes. In this chapter, methods are presented for using the Barley stripe mosaic virus VIGS system (BSMV-VIGS) to silence genes in hexaploid wheat.
病毒诱导的基因沉默(VIGS)是一种有用的功能基因组学工具,可快速创建植物基因敲除表型,用于推断基因功能。直到最近,VIGS仅在双子叶植物中可行。然而,基于大麦条纹花叶病毒(BSMV)的克隆载体的开发现已使VIGS在大麦和小麦中成为可能。VIGS在小麦功能基因组学中具有特殊优势,因为该生物体的六倍体特性和转化抗性极大地阻碍了基因功能鉴定策略。在本章中,介绍了使用大麦条纹花叶病毒VIGS系统(BSMV-VIGS)在六倍体小麦中沉默基因的方法。