In spring 2023, during a survey of Eucalyptus trees (Eucalyptus camaldulensis Dehn.) in Jiroft, Kerman Province, Iran, leaves with leaf spot symptoms were collected. Fungal colonies with similar growth patterns were isolated from symptomatic leaves. The recovered isolates were identified based on the combination of morphological features and sequence data of the ITS-rDNA and part of the translation elongation factor 1-alpha (tef-1a) gene as Pestalotiopsis biciliata. Pathogenicity tests were carried out by using detached leaves and seedlings of Eucalyptus, fulfilling the Koch's postulates. To our knowledge, this is the first report of P. biciliata for the Iranian Funga and also the first report of the pathogenicity of this species on E. camaldulensis in Iran.