Forest trees withering is an increasingly ubiquitous phenomenon throughout the Mediterranean basin. Aleppo pine (Pinus halepensis Mill.) is one of the dominant conifer species in this region. In the northeastern pine coastal woodland of Tunisia, Pinus halepensis stands originally afforested to stabilize sand dunes are often damaged. Fungal pathogens are among the main causes of forest tree diseases. Species of the Botryosphaeriaceae family, having a worldwide distribution, are well known as plant pathogens including Pinus spp. Pinus halepensis trees are suffering from shoot dieback, branch cankers and trunk cankers in two Tunisian forests. The objective of the present work was to characterize Botryosphaeriaceae species associated with P. halepensis branch canker in Bizerte (Rimel forest). A total of 43 isolates were obtained from infected branches of P. halepensis trees. Using morphological characteristics and phylogenetic analysis, two Botryosphaeriaceae species were identified, that is Diplodia scrobiculata and Neofusicoccum luteum. Pathogenicity tests were performed on 3‐year‐old P. halepensis plants and both isolates were found to be pathogenic. Diplodia scrobiculata was the more virulent species had a greater virulence than Neofusicoccum luteum based on the length of vascular necrosis in the wood of the P. halepensis inoculated stem. To our knowledge, this is the first record of Neofusicoccum luteum and Diplodia scrobiculata associated with P. halepensis branch canker and dieback in Tunisia and worldwide.