During a four-year project conducted to identify fungal species associated with grapevine trunk diseases (GTDs) in Hungarian vineyards, two non-sporulating strains isolated from vines exhibiting typical GTD symptoms were identified as Dothiorella omnivora based on their nrDNA ITS and EF1-α sequences. Conidial production of these strains was induced on pine needle medium where production of spermatia has also been observed. Pathogenicity tests confirmed their virulence on potted vines. Dothiorella omnivora is a recently described species of the Botryosphaeriaceae, known to infect hazelnut, ash, walnut, and other woody hosts. This is the first European record of D. omniovora from vines exhibiting GTD symptoms.