Intraspecific diversity of species of the Pseudallescheria boydii complex

In order to establish intraspecific diversity of Pseudallescheria boydii and Scedosporium apiospermum, and to develop tools for their identification, variability within P. boydii and related species was investigated at different levels of diversity. Sequences of the D1/D2 region of large subunit (LSU) and of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) gene were analyzed for a set of 57 strains, as well as partial sequences of the elongation factor 1-alpha (EF 1-alpha). Incongruence among 3 locus lineages was detected by partition homogeneity test (PHT). The maximum parsimony (MP) tree of the combined sequence data set, with the exception of strain CBS 499.90, formed 3 clades with high bootstrap support, corresponding to previously described nuclear DNA (nDNA)/DNA reassociation groups. These groups are known to differ slightly in predilection and temperature relations. Using Structure software, population genetic analysis revealed 3 clusters within the complex on the basis of multi-locus genotype data. Strain distribution in the clusters was concordant with that in the 3 clades of combined multi-locus MP tree. Recombination among individuals of a clade in evolutional history was found in 2 of the 3 clades. There was population differentiation among the 3 clades. Restriction fragment length polymorphism (RFLP) analysis of the intergenic spacer (IGS) region of rDNA gene was added to further characterize subspecific entities. When the IGS regions of 22 strains were digested with the restriction endonucleases Hae III and Mbo I, seven and five distinct patterns were detected, respectively. This subtyping did not reveal any correspondence with geographic origin or clinical appearance. Though we need more evidence to locate the 3 clades of the P. boydii complex at species or population level, the sequence of the D1/D2 region is sufficiently variable for identification of taxa belonging to the P. boydii complex.