Isolation of apoplastic fluid from leaf tissue by the vacuum infiltration-centrifugation technique

Upon infection of plants by pathogens, at least at the early stages of infection, the interaction between the two organisms occurs in the apoplast. To study the molecular basis of host susceptibility vs. resistance on the one hand, and pathogen virulence vs. avirulence on the other, the identification of extracellular compounds such as pathogen effectors that determine the outcome of the interaction is essential. Here, I describe the vacuum infiltration-centrifugation technique, which is an extremely simple and straightforward method to explore one of the most important battlefields of a plant-pathogen interaction; the apoplast.