Effector Proteins of the Bacterial Pathogen Pseudomonas syringae Alter the Extracellular Proteome of the Host Plant, Arabidopsis thaliana

In plants, potential pathogenic bacteria do not enter the host cell. Therefore, a large portion of the molecular interaction between microbial pathogen and host occurs in the extracellular space. To investigate potential mechanisms of disease resistance and susceptibility, we analyzed changes in the extracellular proteome, or secretome, using the Arabidopsis-Pseudomonas syringae pathosystem. This system provides the possibility to directly compare interactions resulting in basal resistance, susceptibility, and gene-specific resistance by using different genotypes of Pseudomonas on the same host. After infecting suspension-cultured cells of Arabidopsis with the Pseudomonas strain of interest, we isolated protein from the cell culture medium representing the secretome. After one-dimensional gel separation and in-gel digestion of proteins, we used iTRAQ (isobaric tags for relative and absolute quantitation) labeling in conjunction with LC-MS/MS to perform relative quantitative comparisons of the secretomes from each of these interactions. We obtained quantitative information from 45 Arabidopsis proteins that were present in all three biological experiments. We observed complex patterns of accumulation, ranging from proteins that decreased in abundance in the presence of all three bacterial strains to proteins that specifically increased or decreased during only one of the interactions. A particularly intriguing result was that the virulent bacteria (e. g. a susceptible interaction) caused the extracellular accumulation of a specific subset of host proteins lacking traditional signal peptides. These results indicate that the pathogen may manipulate host secretion to promote the successful invasion of plants. Molecular & Cellular Proteomics 8:145-156, 2009.