Regulation of Xylella fastidiosa virulence factors by c-di-GMP phosphodiesterases

Xylella fastidiosa is an important bacterial plant pathogen that colonizes the xylem of hundreds of plant species. X. fastidiosa cause Pierce's disease in grapevine by occlusion of the xylem by extensive bacterial colonization, extracellular polysaccharides and the formation of a biofilm. These traits are mediated in a cell-density manner by a cell-to-cell signaling system that transduces a diffusible signaling factor (DSF). This dissertation demonstrates that PD1994, PD1617 and RpfG regulate important traits for bacterial virulence such as cell-cell signaling, biofilm formation and cell aggregation. X. fastidiosa strains harboring mutations in pd1994 (which encodes for a defective GGDEF- EAL-domain protein) and in pd1617 (which encodes for a EAL-domain protein) have increased growth rate, increased biofilm formation, increased plant colonization and decreased cell aggregation. Gene expression analysis of the pd1994 mutant strain showed overexpression of rpfF, which is a DSF synthase, suggesting that PD1994 regulates DSF signaling by repressing rpfF expression. Additionally, the pd1994mutant showed overexpression of pd1617 and rpfG (with EAL and HD-GYP domains respectively, that may be responsible for c-di-GMP turnover), which suggested that this mutant may have low c-di-GMP levels and that PD1994 regulates c-di-GMP turnover by repression of RpfG activity and PD1617 gene expression. X. fastidiosa harboring a mutation on rpfG exhibited decreased biofilm formation while it had no effect in growth or cell aggregation. Together, these results suggest that PD1994, PD1617 and RpfG regulate the DSF regulatory network by controlling the turnover of the second messenger c-di-GMP.