Xylella fastidiosa (Xf) is a xylem-limited bacterial pathogen that causes leaf scorch symptoms in numerous plant species in urban, agricultural, and natural ecosystems worldwide. The exact mechanism of hydraulic disruption and systemic colonization of xylem byXf remains elusive across all host plants. To understand both processes better, the functional and structural characteristics of xylem in different organs of both healthy andXf-infected trees of severalQuercus species were studied. Hydraulic conductivity (K(s)) inXf-infected petioles ofQ. palustris andQ. rubra decreased significantly compared with healthy trees as the season progressed and plummeted to zero with the onset of scorch symptoms. Prior to the onset of symptoms, embolism was as much as 3.7 times higher inXf-infected petioles compared with healthy controls and preceded significant reductions inK(s). Embolism likely resulted from pit membrane degradation during colonization of new petiole xylem and triggered the process that eventually led to vessel occlusion. Pit membrane porosity was studied using the following four methods to determine if a pathway exists in the xylem network of woody stems that allows for passiveXf migration: (i) calculations based on vulnerability to cavitation data, (ii) scanning electron micrographs, (iii) microsphere injections, and (iv) air seeding thresholds on individual vessels. All four methods consistently demonstrated that large pit membrane pores (i.e. greater than the diameter of individualXf) occur frequently throughout the secondary stem xylem in several Quercus species. These large pores probably facilitate systemic colonization of the secondary xylem network and contribute to the high susceptibility to bacterial leaf scorch exhibited among these species.