title: Genome size and ploidy of Paracoccidioides brasiliensis reveals a haploid DNA content: flow cytometry and GP43 sequence analysis.
abstract: The aim of this study was to evaluate genome size and ploidy of the dimorphic pathogenic fungus Paracoccidioides brasiliensis. The cell cycle analysis of 10 P. brasiliensis isolates by flow cytometry (FCM) revealed a genome size ranging from 26.3+/-0.1Mb (26.9+/-0.1fg) to 35.5+/-0.2Mb (36.3+/-0.2fg) per uninucleated yeast cell. The DNA content of conidia from P. brasiliensis ATCC 60855-30.2+/-0.8Mb (30.9+/-0.8fg) -showed no significant differences with the yeast form, possibly excluding the occurrence of ploidy shift during morphogenesis. The ploidy of several P. brasiliensis isolates was assessed by comparing genome sizing by FCM with the previously described average haploid size obtained from electrophoretic karyotyping. The analysis of intra-individual variability of a highly polymorphic P. brasiliensis gene, GP43, indicated that only one allele seems to be present. Overall, the results showed that all analysed isolates presented a haploid, or at least aneuploid, DNA content and no association was detected between genome size/ploidy and the clinical-epidemiological features of the studied isolates. This work provides new knowledge on P. brasiliensis genetics/genomics, important for future research in basic cellular/molecular mechanisms and for the development/design of molecular techniques in this fungus.
authors.affiliation: Life and Health Sciences Research Institute (ICVS), School of Health Sciences, University of Minho, Braga, Portugal.
authors.forename: A J
authors.initials: AJ
authors.lastname: Almeida
authors.forename: D R
authors.initials: DR
authors.lastname: Matute
authors.forename: J A
authors.initials: JA
authors.lastname: Carmona
authors.forename: M
authors.initials: M
authors.lastname: Martins
authors.forename: I
authors.initials: I
authors.lastname: Torres
authors.forename: J G
authors.initials: JG
authors.lastname: McEwen
authors.forename: A
authors.initials: A
authors.lastname: Restrepo
authors.forename: C
authors.initials: C
authors.lastname: Leão
authors.forename: P
authors.initials: P
authors.lastname: Ludovico
authors.forename: F
authors.initials: F
authors.lastname: Rodrigues
mesh_terms: D000946:Antigens, Fungal / Q000235:genetics*; D015825:Chromosomes, Fungal; D004271:DNA, Fungal / Q000032:analysis* / Q000737:chemistry; D005434:Flow Cytometry; D005656:Fungal Proteins / Q000235:genetics*; D016681:Genome, Fungal; D006023:Glycoproteins / Q000235:genetics*; D006238:Haploidy; D007621:Karyotyping; D010228:Paracoccidioides / Q000145:classification / Q000235:genetics* / Q000254:growth & development; D011003:Ploidies; D017422:Sequence Analysis, DNA
chemical_list: C103180:43 kDa protein, Paracoccidioides; D000946:Antigens, Fungal; D004271:DNA, Fungal; D005656:Fungal Proteins; D006023:Glycoproteins