Little is known regarding the fungi, especially fungal pathogens, associated with mangroves in Africa. This includes fungi in the Botryosphaeriaceae that comprise numerous opportunistic, stress-associated pathogens often associated with trees affected by environmental and anthropogenically generated stresses, such as those affecting mangroves. We investigated the occurrence of endophytic Botryosphaeriaceae along the entire distribution of mangroves in South Africa. Asymptomatic branches were collected from ten localities and six mangrove species. Isolates resembling species of Botryosphaeriaceae were identified based on multi-gene sequence data of the internal transcribed spacer regions (ITS), including the 5.8S nrRNA, the beta-tubulin (tub2), partial translation elongation factor 1-alpha (tef1-α), and DNA-directed RNA polymerase II second largest subunit (rpb2) gene regions. Inoculation trials were conducted on healthy branches of Avicennia marina and Bruguiera gymnorrhiza to evaluate the potential pathogenicity of the collected species. Fourteen species in the Botryosphaeriaceae belonging to four genera, Botryosphaeria, Diplodia, Lasiodiplodia, and Neofusicoccum were collected, including five new species. Neofusicoccum was the most prevalent genus followed by Lasiodiplodia, with species of Diplodia and Botryosphaeria being the least frequent. The inoculation studies revealed that one of the new species, Lasiodiplodia avicenniae is highly pathogenic to A. marina and could pose a threat to the health of these trees.