In addition to Eutypa lata, which causes eutypa dieback, numerous other fungi in the Diatrypaceae family have been isolated from diseased grapevines (Vitis vinifera) and other woody hosts. Pathogenicity trials comprising 70 strains of diatrypaceous fungi comprising nine species in six genera were conducted to determine if these fungi, collected in Australia, were pathogenic to grapevines. When inoculated into wounded trunks of cv. Cabernet Sauvignon, eight species, including Eutypa lata, Eutypa leptoplaca, Cryptovalsa ampelina, Cryptovalsa rabenhorstii, Eutypella citricola, Eutypella microtheca, Diatrypella vulgaris and Diatrype sp. produced necrotic lesions significantly longer than on controls. In addition, all nine species (including Cryptosphaeria sp.) were reisolated from the margins of developing lesions, and at varying distances above and below the point of inoculation. Diatrypaceous fungi were frequently isolated from asymptomatic or otherwise healthy tissue several centimeters ahead of the disease margin. Methods to control diseases associated with diatrypaceous fungi must take into account their propensity to colonize woody tissues ahead of, or in the absence of visible symptoms. Current recommendations for the management of eutypa dieback using remedial surgery and pruning wound protection appear sufficient for the control of the other diatrypaceous fungi included in this study.