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.
Key wordsVacuum infiltration-centrifugation technique Apoplast Apoplastic fluid Intercellular washing fluid Plant–pathogen interaction Extracellular space Communication Effector Avirulence Resistance
Wladimir Tameling is thanked for his help with making the pictures and for critical reading of the manuscript. I thank the Centre for BioSystems Genomics (CBSG), which is part of the Netherlands Genomics Initiative and NWO, for support.
- 3.Kaschani F, Shabab M, Bozkurt T, Shindo T, Schornack S, Gu C, Ilyas M, Kamoun S, Van der Hoorn RAL (2010) An effector-targeted protease contributes to defense against Phytophthora infestans and is under diversifying selection in natural hosts. Plant Physiol 154: 1794–1804.PubMedCrossRefGoogle Scholar
- 10.Rooney HCE, Van ’t Klooster JW, Van der Hoorn RAL, Joosten MHAJ, Jones JDG, De Wit PJGM (2005) Cladosporium Avr2 inhibits tomato Rcr3 protease required for Cf-2-dependent disease resistance. Science 308: 1783–1786.Google Scholar