Mei Yu, Whitham Steven A
Department of Plant Pathology and Microbiology, Iowa State University, 351 Bessey, Ames, IA, 50011, USA.
Methods Mol Biol. 2018;1676:129-139. doi: 10.1007/978-1-4939-7315-6_7.
Virus-induced gene silencing (VIGS) is a powerful technology for rapidly and transiently knocking down the expression of plant genes to study their functions. A VIGS vector for maize derived from Foxtail mosaic virus (FoMV), a positive-sense single-stranded RNA virus, was recently developed. A cloning site created near the 3' end of the FoMV genome enables insertion of 200-400 nucleotide fragments of maize genes targeted for silencing. The recombinant FoMV clones are inoculated into leaves of maize seedlings by biolistic particle delivery, and silencing is typically observed within 2 weeks after inoculation. This chapter provides a protocol for constructing FoMV VIGS clones and inoculating them into maize seedlings.
病毒诱导的基因沉默（VIGS）是一种强大的技术，可快速、短暂地抑制植物基因的表达以研究其功能。最近开发了一种源自狐尾花叶病毒（FoMV，一种正义单链RNA病毒）的玉米VIGS载体。在FoMV基因组3'端附近创建的克隆位点能够插入靶向沉默的玉米基因的200 - 400个核苷酸片段。通过生物弹道粒子递送将重组FoMV克隆接种到玉米幼苗的叶片中，接种后通常在2周内观察到基因沉默。本章提供了构建FoMV VIGS克隆并将其接种到玉米幼苗中的方案。
