We reported seven compounds isolated from mycelial cultures of Chilean Basidiomycetes. The compound structures were elucidated by spectroscopic methods. Two polyacetylenes were isolated from Gymnopilus spectabilis: hepta-4,6-diyn-3-ol(1) and 7-chloro-hepta-4,6-diyn-3 ol (2). We isolated the aromatic compounds p-anisaldehyde (3) and 3-chloro-p-anisaldehyde (4) from Hypholoma capnoides. H. fasciculare produced the compounds (3), (4),3,5-dichloro-4-metoxi-benzyl alcohol (5) and the sesquiterpen naematolon (6). Finally, H. sublateritiumproduced (5) and another sesquiterpen, marasmal (7). The compounds (1), (2), (5), and (7) showed biological activity against the tested model microorganism