Plant Pathology and Entomology, NIAB, Cambridge, UK.
Biointeractions and Crop Protection, Rothamsted Research, Harpenden, UK.
Methods Mol Biol. 2022;2523:93-112. doi: 10.1007/978-1-0716-2449-4_7.
One of the important armories that pathogens utilize to successfully colonize the plants is small secreted effector proteins, which could perform a variety of functions from suppression of plant innate immunity to manipulation of plant physiology in favor of the disease. Plants, on the other hand, evolved disease resistance genes that recognize some of the effectors or avirulence (Avr) proteins. Both, identification of the Avr proteins and understanding of the mechanisms of action of other effectors, are important areas of research in the molecular plant-pathogen interactions field as this knowledge is critical for the development of new effective pathogen control measures. To enable functional analysis of the effectors, it is desirable to be able to overexpress them readily in the host plants. Here we describe detailed experimental protocols for transient effector overexpression in wheat and other monocots using binary Barley stripe mosaic virus (BSMV)- and Foxtail mosaic virus (FoMV)-derived vectors. This functional genomics tool, better known as VOX (Virus-mediated protein OvereXpression), is rapid and relatively simple and inexpensive.
病原体用来成功定殖植物的重要武器之一是小分泌效应子蛋白，这些蛋白可以执行多种功能，从抑制植物先天免疫到操纵植物生理学以利于疾病。另一方面，植物进化出了识别一些效应子或无毒（Avr）蛋白的抗病基因。鉴定 Avr 蛋白和了解其他效应子的作用机制都是分子植物-病原体相互作用领域的重要研究领域，因为这些知识对于开发新的有效病原体控制措施至关重要。为了能够对效应子进行功能分析，理想情况下能够在宿主植物中容易地过表达它们。在这里，我们描述了使用二元大麦黄花叶病毒（BSMV）和谷子花叶病毒（FoMV）衍生载体在小麦和其他单子叶植物中瞬时过表达效应子的详细实验方案。这种功能基因组学工具，通常称为 VOX（病毒介导的蛋白过表达），快速、相对简单且廉价。
