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Forty fungi isolated from diverse environments in Western Australia were screened for ligninolytic activity based on in-vivo decolorization of the polymeric dye Poly R-478. Three isolates identified as Aspergillus, Botryosphaeria and Coniochaeta species were selected for further studies. The Botryosphaeria and Coniochaeta isolates were found to produce laccase constitutively in submerged culture when grown on glucose, or on ryegrass seed by solid state fermentation. A comparison of the three isolates grown on glucose in the presence of 3,4-dimethoxybenzyl (veratryl) alcohol (40 mmol l-1) showed that only the Botryosphaeria isolate produced laccase, with laccase activities 115-fold higher than when grown on glucose alone.