Wood-rot fungi have been shown to be powerful agents in many biotechnological processes. However, there may be great inter- and intra-specific variability in their performance. Consequently, it is not only important to screen a broad range of species but also different isolates of the same species in order to obtain strains with specific biotechnological profiles. In this study, the presence of wood-rot fungi was monitored in Southern-Central Chile and the biotechnological potential of the isolates was analyzed by determining lignocellulolytic enzymes and tolerance to metal ions (Cu and Cd) in solid medium. Seventy-one strains were isolated from cultures of a total of 144 basidiomes collected from wood substrates and 59 species of 18 different genera were identified, of which four are first records for Chile: Antrodia xantha, Gloeophyllum abietinum, G. protractum and Stereum rameale. Cellulase and xylanase activity were detected in all strains and 20 strains showed significant ligninolytic activity. The great majority of the strains showed tolerance to 3mM Cu in solid medium, but were inhibited by 1mM Cd. In contrast, some strains of the white-rot fungi Ganoderma australe, Stereum hirsutum and Trametes versicolor presented high lignocellulolytic potential combined with metal tolerance. Possible applications of these strains in biodegradation or bioremediation processes are discussed.3611860868