A large collection of Botryosphaeriaceae isolates obtained from fruit and forest trees with fruit rot, canker and dieback disease symptoms in northern provinces of the country were examined in this study. Based on morphology and sequence data (ITS and EF1-α), two species, Diplodia mutila and Spencermartinsia viticola are illustrated and described as new records for Iran mycobiota. Furthermore, D. sapinea morphotype A is determined here for the first time from Iran