Actinomycetes, one of the major communities of the microbial population present in soil, can also be found inhabiting a wide diversity of ecological sources. We have explored the use of lichens as an alternative source for the isolation of novel actinomycetes. Lichens are symbiotic mixtures of fungi, green algae and/or cyanobacteria and whereas these symbiotic components have been extensively described, the microbial community inhabiting this niche has not been well characterized. We studied the diversity of the actinomycete population isolated from lichens collected in tropical areas from the Hawaii and Reunion islands and in cold areas from Alaska. The diversity of the microbial population was evaluated using fatty acid analysis and molecular fingerprinting. A PCR approach to screen the isolates for genes associated with secondary metabolite production was applied to evaluate the biosynthetic potential of these strains; profiles obtained for each isolate were compared to the antimicrobial activity exhibited by these isolates in laboratory conditions. Our results demonstrate that lichens represent an extremely rich reservoir for the isolation of a wide diversity of actinomycetes many of them representing still today a rich untapped source of secondary metabolites.