Vaghchhipawala Zarir, Rojas Clemencia M, Senthil-Kumar Muthappa, Mysore Kirankumar S
Plant Biology Division, The Samuel Roberts Noble Foundation, Ardmore, OK, USA.
Methods Mol Biol. 2011;678:65-76. doi: 10.1007/978-1-60761-682-5_6.
Agroinoculation, first developed as a simple tool to study plant-virus interactions, is a popular method of choice for functional gene analysis of viral genomes. With the explosive growth of genomic information and the development of advanced vectors to dissect plant gene function, this reliable method of viral gene delivery in plants, has been recruited and morphed into a technique popularly known as agroinfiltration. This technique was developed to examine the effects of transient gene expression, with applications ranging from studies of plant-pathogen interactions, abiotic stresses, a variety of transient expression assays to study protein localization, and protein-protein interactions. We present a brief overview of literature which document both these applications, and then provide simple agroinoculation and agroinfiltration methods being used in our laboratory for functional gene analysis, as well as for fast-forward and reverse genetic screens using virus-induced gene silencing (VIGS).
农杆菌接种最初是作为研究植物-病毒相互作用的一种简单工具而开发的,是病毒基因组功能基因分析的一种常用方法。随着基因组信息的爆炸式增长以及用于剖析植物基因功能的先进载体的发展,这种在植物中可靠的病毒基因传递方法已被采用并演变成一种广为人知的技术,即农杆菌浸润法。该技术是为研究瞬时基因表达的影响而开发的,其应用范围从植物-病原体相互作用、非生物胁迫的研究,到用于研究蛋白质定位和蛋白质-蛋白质相互作用的各种瞬时表达分析。我们简要概述了记录这两种应用的文献,然后提供了我们实验室用于功能基因分析以及使用病毒诱导基因沉默(VIGS)进行正向和反向遗传筛选的简单农杆菌接种和农杆菌浸润方法。