细菌细胞外囊泡(BEV)从细菌细胞的表面释放并且含有不同的分子货物。主要与哺乳动物的细菌病原体进行的研究表明,BEV参与多个过程,如细胞-细胞通讯,RNA的传递,DNA,和蛋白质靶向细胞,保护免受压力,操纵宿主免疫力,和其他功能。直到十年前,BEV在植物-细菌相互作用中的作用几乎没有研究。然而,最近的研究表明,植物病原体的BEV具有与哺乳动物病原体相似的功能,现在更多的研究致力于研究它们与植物的作用和相互作用。在以下方法一章中,我们提供了5种经过充分验证的检测方法来检测BEV与植物免疫系统的相互作用.这些检测依赖于不同的标记或免疫输出,这表明植物免疫的激活(防御标记基因表达,活性氧爆发,幼苗抑制)。此外,我们提供的检测方法可以直接评估BEV攻击后免疫系统的启动情况及其对随后的局部或全身感染的反应的有效性.总之,这些分析为BEV和植物免疫系统的相互作用提供了彻底的检查。
Bacterial extracellular vesicles (BEVs) are released from the surface of bacterial cells and contain a diverse molecular cargo. Studies conducted primarily with bacterial pathogens of mammals have shown that BEVs are involved in multiple processes such as cell-cell communication, the delivery of RNA, DNA, and proteins to target cells, protection from stresses, manipulation of host immunity, and other functions. Until a decade ago, the roles of BEVs in plant-bacteria interactions were barely investigated. However, recent studies have shown that BEVs of plant pathogens possess similar functions as their mammalian pathogen counterparts, and more research is now devoted to study their roles and interactions with plants. In the following methods chapter, we provide five well-validated assays to examine the interaction of BEVs with the plant immune system. These assays rely on different markers or immune outputs, which indicate the activation of plant immunity (defense marker gene expression, reactive oxygen species burst, seedling inhibition). Furthermore, we offer assays that directly evaluate the priming of the immune system following BEV challenge and the effectiveness of its response to subsequent local or systemic infection. Altogether, these assays provide a thorough examination to the interactions of BEVs and the plant immune system.