The effects of carrier DNAs on the transformation of the basidiomycete Pleurotus ostreatus were analyzed. When lambda phage DNA was added to a transformation mixture containing protoplasts and Cbx(R) vector plasmid, an increased number of drug-resistant transformants was observed on a screening plate containing 2 mug carboxin/ml. The highest efficiency (about 200 transformants/mug Vector plasmid) was obtained by the addition of heat-denatured lambda DNA, which gave yields approximately 50-fold higher than the control experiment without a carrier DNA. To our knowledge, this is the first report on enhancement in transformation efficiency of fungal protoplasts by single strand carrier DNA.