Department of Plant Pathology and Weed Research, Agricultural Research Organization - Volcani Institute, Rishon LeZion, Israel.
Methods Mol Biol. 2024;2843:95-117. doi: 10.1007/978-1-0716-4055-5_7.
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.
细菌细胞外囊泡 (BEVs) 从细菌细胞表面释放出来，包含多种分子货物。主要针对哺乳动物细菌病原体进行的研究表明，BEVs 参与多种过程，如细胞间通讯、将 RNA、DNA 和蛋白质递送至靶细胞、抵抗应激、操纵宿主免疫等功能。直到十年前，BEVs 在植物-细菌相互作用中的作用几乎没有被研究过。然而，最近的研究表明，植物病原体的 BEVs 具有与其哺乳动物病原体对应物相似的功能，现在更多的研究致力于研究它们的作用及其与植物的相互作用。在以下方法章节中，我们提供了五种经过充分验证的测定法来研究 BEVs 与植物免疫系统的相互作用。这些测定法依赖于不同的标记物或免疫输出，这些输出指示植物免疫的激活（防御标记基因表达、活性氧爆发、幼苗抑制）。此外，我们提供了直接评估 BEV 挑战后免疫系统的启动及其对随后的局部或系统感染的反应有效性的测定法。总的来说，这些测定法提供了对 BEVs 和植物免疫系统相互作用的全面检查。
