In this study, we undertook a functional characterization and transcriptome analysis that enabled a comprehensive study of the mating type loci of the mushroom Schizophyllum commune. Induced expression of both the bar2 receptor and the bap2(2) pheromone gene within 6 to 12 h after mates' contact was demonstrated by quantitative real-time PCR. Similar temporal expression patterns were confirmed for the allelic bbr1 receptor and bbp1 pheromone-encoding genes by Northern hybridization. Interestingly, the fusion of clamp connections to the subterminal cell was delayed in mating interactions in which one of the compatible partners expressed the bar2 receptor with a truncated C terminus. This developmental delay allowed the visualization of a green fluorescent protein (Gfp)-labeled truncated receptor at the cell periphery, consistent with a localization in the plasma membrane of unfused pseudoclamps. This finding does not support hypotheses envisioning a receptor localization to the nuclear membrane facilitating recognition between the two different nuclei present in each dikaryotic cell. Rather, Gfp fluorescence observed in such pseudoclamps indicated a role of receptor-pheromone interaction in clamp fusion. Transcriptome changes associated with mating interactions were analyzed in order to identify a role for pheromone-receptor interactions. We detected a total of 89 genes that were transcriptionally regulated in a mating type locus A-dependent manner, employing a cutoff of 5-fold changes in transcript abundance. Upregulation in cell cycle-related genes and downregulation of genes involved in metabolism were seen with this set of experiments. In contrast, mating type locus B-dependent transcriptome changes were observed in 208 genes, with a specific impact on genes related to cell wall and membrane metabolism, stress response, and the redox status of the cell.T he filamentous fungus Schizophyllum commune is a widely distributed mushroom that lives saprophytically and can cause white rot on wood. It has been extensively used to study mating interactions for almost 100 years (47). In addition to studies involving the mushroom Coprinopsis cinerea and the corn smut Ustilago maydis, the mating type genes have been extensively analyzed for this tetrapolar basidiomycete (4, 29). The recent publication of the S. commune genome sequence enables global expression analyses (41). During the life cycle of this fungus, compatible haploid monokaryons can mate to produce a fertile dikaryon with two nuclei per cell, one derived from each mating partner. Under appropriate environmental conditions, the dikaryon is able to form the sexual reproductive fruiting bodies (40). While the fusion of two monokaryotic hyphae (plasmogamy) is independent of mating types, specific steps of sexual development are regulated by the mating type genes encoded in the loci A and B (25, 47). For each of these loci, two linked, multiallelic subloci (termed ␣ and ␤) have been determined by recombination analyses (27,49,51,65). The A loci encode homeodomain ...

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