First Report of Diplodia fraxini and Diplodia subglobosa Causing Canker and Dieback of Fraxinus excelsior in Slovenia

In recent decades the vitality and productivity of European ash trees in Slovenia have been reduced by the onset of canker and dieback disease symptoms on young and old trees, identified primarily as ash dieback caused by Hymenoscyphus fraxineus. Given the limited information available about the etiology of this emerging disease, a study was carried out to isolate, identify, and characterize the fungal species involved in the observed ash symptoms. Field surveys were conducted in five forest sites where 50 symptomatic branch samples were collected. All samples were inspected and used for fungal isolation. Based on morphology, colony appearance, and DNA sequence data of the internal transcribed spacer region, 125 fungal colonies belonging to five species were isolated and identified. Only a few symptomatic ash samples yielded colonies of H. fraxineus, whereas Botryosphaeriaceae species were isolated with a high frequency, with Diplodia fraxini as the dominant species. A pathogenicity test proved that all isolated species were pathogenic on European ash, causing bark lesions and wood discoloration. All Botryosphaeriaceae species isolated in this study are reported for the first time on European ash in Slovenia.