Pitch canker, caused by Fusarium circinatum, is a destructive disease of Pinus species and has recently been shown to represent a substantial threat to natural and commercial forests in northern Spain. The genetic diversity of F. circinatum in the Basque Country of Spain was assessed by characterising 96 isolates based on vegetative compatibility groups (VCGs), mating type assays, polymorphic DNA-markers and amplified fragment length polymorphism (AFLP) analyses. For this purpose, F. circinatum isolates were collected from diseased Pinus radiata as well as from insects associated with this host. Overall, a low level of diversity was detected in the population. The isolates represented only two VCGs and they were all of the same mating type. AFLP analyses revealed three genotypes and polymorphic DNA-markers specific for F. circinatum showed nine genotypes. The most common genotypes represented 97 % of all isolates for AFLP analysis and 68 % of isolates for the polymorphic DNA-marker sets. Over all, this indicates that pitch canker in the Basque Country of Spain is caused by a clonally propagating population of F. circinatum, typical of a recently introduced pathogen.