Zhou Bangjun, Zeng Lirong
Department of Plant Pathology, Center for Plant Science Innovation, University of Nebraska, Lincoln, NE, USA.
Southern Regional Collaborative Innovation Center for Grain and Oil Crops, Hunan Agricultural University, Changsha, China.
Methods Mol Biol. 2018;1743:51-63. doi: 10.1007/978-1-4939-7668-3_5.
Plants have evolved a sophisticated innate immune system to contend with potential infection by various pathogens. Understanding and manipulation of key molecular mechanisms that plants use to defend against various pathogens are critical for developing novel strategies in plant disease control. In plants, resistance to attempted pathogen infection is often associated with hypersensitive response (HR), a form of rapid programmed cell death (PCD) at the site of attempted pathogen invasion. In this chapter, we describe a method for rapid identification of genes that are essential for plant innate immunity. It combines virus-induced gene silencing (VIGS), a tool that is suitable for studying gene function in high-throughput, with the utilization of immunity-associated PCD, particularly HR-linked PCD as the readout of changes in plant innate immunity. The chapter covers from the design of gene fragment for VIGS, the agroinfiltration of the Nicotiana benthamian plants, to the use of immunity-associated PCD induced by twelve elicitors as the indicator of activation of plant immunity.
植物已经进化出一套复杂的先天免疫系统来应对各种病原体的潜在感染。理解和操纵植物用于抵御各种病原体的关键分子机制对于开发植物病害控制的新策略至关重要。在植物中,对病原体侵染企图的抗性通常与超敏反应(HR)相关,超敏反应是在病原体入侵企图部位发生的一种快速程序性细胞死亡(PCD)形式。在本章中,我们描述了一种快速鉴定植物先天免疫所必需基因的方法。它将病毒诱导的基因沉默(VIGS)(一种适用于高通量研究基因功能的工具)与利用免疫相关的PCD(特别是与HR相关的PCD)相结合,作为植物先天免疫变化的读数。本章内容涵盖从用于VIGS的基因片段设计、本氏烟草植株的农杆菌浸润,到使用由十二种激发子诱导的免疫相关PCD作为植物免疫激活指标的过程。