A molecular survey of TYLCV/TYLCSV and their associated vector Bemisia tabaci, was performed during 2004-2005 in five regions of Southern Italy: i.e. Sardinia (one locations), Sicily (one location), Calabria (three locations), Campania (two locations) and Basilicata (one location). A total of 71 tomato samples were checked for virus infection and for the presence of the vector. Degenerate primers allowing the amplification of the coat protein gene of both TYLCSV and TYLCV isolates were designed. PCR fragments were then digested with restriction endonuclease Ava II, which was expected to cut TYLCSV differently from TYLCV. Results clearly suggested that in all the inspected Italian regions the two viruses are widespread and present in single plant both alone and in mixed infections. The identity of the two viruses was confirmed by total or partial sequencing of field isolates. Concerning the populations of the B. tabaci associated with TYLCD epidemics, the molecular characterization of COI gene (citocrome oxidase I) indicated that Q biotype was the most prevalent biotype. This fact might be the result of the large use of some insecticides against which Q biotype populations easily develop resistances, as already confirmed in some countries of Mediterranean basin.