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Abstract: The edible mushrooms Agrocybe aegerita, Agaricus bisporus, Lentinus edodes, Pleurotus ostreatus and Volvariella volvacea have now been transformed; the first step towards directed genetic modification of these species. In A. bisporus, we have evaluated a range of vectors, delivery systems, and Agrobacterium transfection. Using protoplasts and selection for hygromycin resistance some transformants were obtained, but they proved unstable. Agrotransfections did yield stable transformants. To improve efficiency and utility of markers we are developing novel systems based on Agaricus genes and regulatory sequences. An extensive range of promoter cassettes has been built for use with a variety of reporter/marker genes and to assist development of future constmcts. The A. bisporus orotidine-5'-monophosphatedecarboxylase gene (URA3) which has potential for transformation of a ura3 auxotroph, was cloned and sequenced. Genes forpara-aminobenzoic acid can encode sulphonamide resistance and this system was tested in homobasidiomycetes. A mutants, bisporus gene that encodes carboxin resistance was cloned, sequenced and used to transform Coprinus cinereus.
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