Unveiling the mating system and genetic variability in the yeast Kwoniella mangroviensis using molecular approaches

Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina

In fungi, sexual reproduction is orchestrated by genomic regions known as mating type loci (MAT), which can be defined by two physically unlinked sex determining regions (tetrapolar mating system) or a single locus (bipolar system). Kwoniella mangroviensis is a saprobic basidiomycetous yeast, belonging to the Kwoniella clade of the order Tremellales, which was described as possessing a bipolar mating-system, similar to the related pathogenic species Cryptococcus neoformans and Cryptococcus gattii of the sister Filobasidiella clade. Studies aimed at elucidating the evolution of the MAT locus of these two Cryptococcus species of clinical importance, targeted several related saprobic species in the Kwoniella and Filobasidiella clades. An evolutionary model ensuing from those studies suggests that the tetrapolar mating-systems found in most species are ancestral and gave rise to the bipolar mating-system of C. neoformans by chromosomal rearrangements and fusion events. The present study comprised strains from the original work describing K. mangroviensis, as well as additional isolates from plant substrates in Europe and Africa. A multilocus sequence typing approach revealed genetic variability among those strains and led to the identification of two novel species closely related to K. mangroviensis: Kwoniella sp. A and Kwoniella sp. B. The mating system of K. mangroviensis and sibling species was further explored by a genetic approach based on sequencing of two MAT genes: STE20 and the divergently transcribed genes SXI1 and SXI2. The results obtained demonstrated tetrapolar mating systems in K. mangroviensis, as well as in Kwoniella sp. A and Kwoniella sp. B. Additionally, the MAT locus structure of K. mangroviensis was unveiled by sequencing a 43 kb genomic region containing the STE20 gene. Twelve genes also present in the MAT loci of related species were identified, and full synteny was found between K. mangroviensis and Cryptococcus heveanensis, a distant member of the Kwoniella clade. These findings provided novel insights into the evolution of MAT loci in basidiomycetous yeasts in the Tremellales.

Fundação para a Ciência e Tecnologia - PTDC/BIA-MIC/113051/2009