Anthracnose is one of the major fungal diseases of strawberry occurring worldwide. In Israel, the disease is caused primarily by the species Colletotrichum acutatum Simmonds. The pathogen causes black spot on fruit, root necrosis and crown rot resulting in mortality of transplants in the field. In addition, lesions are formed on petioles and stolons thus, limiting production of strawberry plants in the nurseries. It is common to find a single botanical species of Colletotrichum infecting multiple hosts, while several Colletotrichum species or biotypes can be associated with a single host. The host range and specificity of C. acutatum from strawberry was examined with pepper, eggplant, tomato, bean and strawberry under greenhouse conditions. The pathogen survived and proliferated in these plants over a three month period without causing any symptoms, except on strawberry. Colonization of the pathogen in these plant tissues was confirmed by a diagnostic test based on PCR-specific primers for C. acutatum. Inoculation of various fruits with C. acutatum from strawberry produced disease symptoms, indicating the potential for cross-infection.