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The aim of this study was to contribute to the knowledge of the richness and distribution of the lichens in the Bío-Bío Region. A total of 120 samples were collected in the surroundings of Termas de Chillán and Las Trancas, Bio-Bio region. The sampling was realized at random considering the high diversity of available substrates. Taxonomical identification was performed on the basis of the analysis of morphological, reproductive and chromatographic (TLC) characters. A total of 41 species were identified, which include 18 families and 31 genera. Out of the total of identified species, 49 % are corticolous, 34 % saxicolous and 17 % terricolous. Of the total of species found, four are new records for Chile: Menegazzia confusa P. James, M. pertransita (Stirt.) R. Sant., Rinodina sophodes (Ach.) A. Massal. and Usnea sinensis Motyka. For each species, information on lichen name, family, substratum, altitude, global distribution, and sampling site is provided. Also, keys to identify the genera found in the study area are included. Pictures for the new records of lichens for the country are also presented. The work contributes to extend the knowledge on the taxa’s distribution in the region. The results suggest that the lichenological expeditions should be continued in unexplored localities of Chile in order to increase the knowledge of the richness, ecology and distribution of lichen biodiversity.