In the past 10 years, the winegrape industry in the southeastern United States has experienced rapid growth; however, further expansion may be inhibited by Pierce's disease (PD). Epidemiological studies were conducted to identify the primary vectors of Xylella fastidiosa, the cause of PD of grape, by surveying sharpshooter population dynamics in the eastern Piedmont and Coastal Plain regions of North Carolina. Sharpshooter species were assessed for the presence of X. fastidiosa in the field. Leafhoppers were trapped in three vineyards in the eastern Piedmont and one vineyard in the northeastern Coastal Plain in 2004 and 2005. Four insects were identified as most abundant: Oncometopia orbona, Graphocephala versuta, Paraphlepsius irroratus, and Agalliota constricta. Adult specimens of O. orbona, G. versuta, and P. irroratus were tested for the presence of X. fastidiosa by nested polymerase chain reaction. In all, 27% of O. orbona, 28% of G. versuta, and 33% of P. irroratus trapped were positive for X. fastidiosa over the two seasons. Transmission experiments demonstrated that both O. orbona and G. versuta have the ability to transmit X. fastidiosa to grape. These vectors are likely to be important in all winegrowing regions of the Southeast, because their presence has been documented throughout the southern states. In DNA analyses, X. fastidiosa strains from insects trapped in North Carolina were genetically similar to one another and to the known “PD strain” from California. This is the first report of these two leafhopper species transmitting X. fastidiosa to grapevines in the Southeast.