Rice blast disease, caused by the fungus Pyricularia grisea (Cooke) Sacc., is responsible for considerable damages in rice crops in Italy and in other parts of the world. This study was conducted in order to investigate the genetic structure of a P. grisea population in the Po area, the largest rice area in Italy. Rice leaves showing blast symptoms were collected in three successive years (1998–2000) and 43 P. grisea monoconidial culture samples were isolated from infected rice leaves. Fungal DNAs were obtained from mycelia. Moreover, six additional P. grisea DNA samples representative for the five characterized European lineages were also investigated. All 49 DNAs were fingerprinted using the Pot2-based repetitive polymerase chain reaction specific for the blast pathogen. Unweighted pair-group method with arithmetic averages cluster analysis shows the presence of three main Italian lineages. Within lineages, similarity was higher than 80%. Samples representative of the three of five known European lineages grouped within these three Italian lineages confirming the presence of three European lineages in Italy. Furthermore, cluster analysis shows the presence of two new haplotypes never found before in the Italian lineage.