Almond anthracnose, caused by Colletotrichum spp. has become an increasing threat in modern almond orchards in Portugal. This study aimed to characterize the genetic, phylogenetic, morphological and virulence diversity of Colletotrichum species associated with symptomatic and asymptomatic almond tree organs collected in the main producing regions of Portugal. A total of 112 isolates were obtained from fruits, leaves, flowers and branches, with most samples originating from the Alentejo region. A