Trees in the Garden Route National Park (GRNP) indigenous forests in South Africa are selectively harvested for timber based on criteria that include signs and symptoms induced by wood-rotting fungi. However, virtually nothing is known regarding the identity and host associations of these macro-fungi in this natural ecosystem. Surveys were conducted in three harvesting compartments in the GRNP to investigate the taxonomic affiliation and species richness of these fungi on standing and recently harvested trees. Samples were collected from basidiomes on infected trees and tree stumps, and from diseased tissues on symptomatic trees. Phylogenetic analyses using ITS sequences characterized the isolates obtained into 26 Operational Taxonomic Units (OTUs) belonging to 17 genera after clustering the sequences at a 97% identity threshold. Ganoderma (Ganodermataceae) and Inonotus (Hymenochaetaceae) were the most species-rich genera and the Bloukrans compartment, with 22 OTUs, showed the highest species richness. A fungus (OTU1) affiliated with Ganoderma pfeifferi was the most abundant in the surveyed areas. Its predominance was also evidenced on host trees since it occurred on 15 of the 20 tree species sampled, with Olea capensis subsp. macrocarpa (Oleaceae) being the most colonized host. Given the wide variety of wood-rotting basidiomycetes revealed by this study and particularly the preponderance of species with pathogenic potential, more attention should be given to better understand their ecological role in this natural ecosystem as well as the effects of logging that may enhance their dissemination or negatively affect their diversity and the health of trees in the region. Fig. 2. Symptoms and signs observed on sampled trees in the GRNP: (a) sap exudation and basal stem rot, (b) bracket-like basidiomes of a Ganoderma species, (c) white mycelial mat under the bark of tree with basal stem rot. [Colour figure can be viewed at wileyonlinelibrary.com]