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Abstract: Wild relatives of the cultivated mushroom, Agaricus bisporus represent an underutilised resource. Knowledge of their breeding biology and relatedness will support their further exploitation as cultivable species and as sources of useful genes. We have used classical and molecular methods to smdy these aspects in six Agaricus species, A. bisporus, A. bitorquis, A. nivescens, whose breeding systems are known, and in A. campestris, A. subfloccosus and A.orvensis whose breeding systems are not well understood and 12 other collections, grouped with A. arvensis by morphological similarity. Breeding systems were classified on the basis of (i) mating interactions with heterokaryosis confirmed by RAPD markers and (ii) the heterogeneity of single spore progeny revealed by RAPD analysis A. campestris was shown to be heterothallic and A. subfloccosus confirmed as homothallic. A. arvensis is heterothallic but homokaryons can frait. RAPD profiles in this species are less heterogeneous than the known heterothallics, A. bitorquis and A. nivescens. ITS sequencing was used to confirm identity and to discriminate between collections in the 'A. arvensis' group. Molecular approaches can, therefore, significantly improve our ability to determine breeding systems and relatedness in the genus Agaricus.
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