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The Indian laurel-leaf fig (<i>Ficus microcarpa</i>) is an important ornamental tree widely distributed in the urban areas of Italy. Surveys conducted in 2019 and 2020 on several tree-lined streets, squares, and public parks in Catania and Siracusa provinces (Sicily, southern Italy) revealed the presence of a new disease on mature trees. About 9% of approximately 450 mature plants showed extensive branch cankers and dieback. Isolations from woody tissues obtained from ten symptomatic plants consistently yielded species belonging to the <i>Botryosphaeriaceae</i> family. The identification of the recovered fungal isolates was based on a multi-loci phylogenetic (maximum parsimony and maximum likelihood) approach of the ITS, <i>tef1</i>-α, and <i>tub2</i> gene regions. The results of the analyses confirmed the presence of three species: <i>Botryosphaeria dothidea</i>, <i>Neofusicoccum mediterraneum</i>, and <i>N. parvum</i>. Pathogenicity tests were conducted on potted, healthy, 4-year-old trees using the mycelial plug technique. The inoculation experiments revealed that all the <i>Botryosphaeriaceae</i> species identified in this study were pathogenic to this host. Previous studies conducted in California showed similar disease caused by <i>Botryosphaeriaceae</i> spp., and the pathogenic role of these fungi was demonstrated. To our knowledge, this is the first report of <i>Botryosphaeriaceae</i> affecting <i>Ficus microcarpa</i> in Europe.