Potato virus Y (PVY), type-member of potyvirus genus, infects a wide host range including cultivated Solanaceous species. This plant virus is worldwide distributed and is the most damaging virus affecting tobacco crops. According to the diversity of biological, serological and molecular caracteristics of PVY, its isolates have been classified into strains, groups and variants. Four host-dependant classifications have been established. Unlike PVY potato strains (classified in groups, PVY^O, PVY^C and PVY^N, and in variants, PVY; and PVY^N-W), tobacco strains were little characterized. The objective is to assess the biological, serological, and molecular diversity of tobacco isolates of PVY in France. A limited range of 21 PVY isolates was characterized. At the biological level, six pathotypes have been described including two new necrotic ones. Serologically, the serotype of most of the tobacco isolates of PVY is O or N. However, 2.6% of the samples analysed were not detected by the monoclonal antibodies initially developed for the potato strains. At the molecular level, the genome of the 21 tobacco isolates of PVY presents some PVY;-like or PVY^N-W-like sequences. These data, showing a great variability of tobacco isolates of PVY, are based however on a limited range of isolates. A large number of samples were collected in 2007 in France. The characterization of some of these isolates will allow to better assess the diversity of biological, serological, and molecular, properties of PVY tobacco isolates.