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Kwoniella mangrovensis has been described as a sexual species with a bipolar mating system. Phylogenetic analysis of multiple genes places this species together withKwoniella heveanensis in theKwoniella clade, a sister clade to that containing two pathogenic species of global importance,Cryptococcus neoformans andCryptococcus gattii , within the Tremellales. Recent studies defining the mating type loci (MAT ) of species in these clades showed that, with the exception ofC. neoformans andC. gattii , which are bipolar with a single biallelic multigeneMAT locus, several other species feature a tetrapolar mating system with two unlinked loci (homeodomain [HD] and pheromone/receptor [P/R] loci). We characterized several strains from the original study describingK. mangrovensis ; twoMAT regions were amplified and sequenced: theSTE20 gene (P/R locus) and the divergently transcribedSXI1 andSXI2 genes (HD locus). We identified five different mating types with differentSTE20 /SXI allele combinations that together with results of mating experiments demonstrate thatK. mangrovensis is not bipolar but instead has a tetrapolar mating system. Sequence and gene analysis for a 43-kb segment of theK. mangrovensis type strainMAT locus revealed remarkable synteny with the homologousK. heveanensis MAT P/R region, providing new insights into slower evolution ofMAT loci in theKwoniella compared to theCryptococcus clade of the Tremellales. The study of additional isolates from plant substrates in Europe and Botswana using a combination of multilocus sequencing withMAT gene analysis revealed two novel sibling species that we nameKwoniella europaea andKwoniella botswanensis and which appear to also have tetrapolar mating systems.

Molecular and Genetic Evidence for a Tetrapolar Mating System in the Basidiomycetous Yeast Kwoniella mangrovensis and Two Novel Sibling Species