| Congresso Brasileiro de Microbiologia 2023 | Resumo: 1246-1 | ||||
Resumo:Paracoccidioides species, as thermodimorphic fungi, are responsible for causing paracoccidioidomycosis (PCM), a severe systemic disease prevalent among individuals exposed to soil-disturbing activities in Latin America. Despite undergoing recent extensive taxonomic reviews due to genetic diversity, cases are commonly attributed to species within the Paracoccidioides brasiliensis complex (P. brasiliensis s. str., P. americana, P. restrepiensis, and P. venezuelensis) or the phylogenetically distinct P. lutzii. We employed a high-throughput mining strategy to enhance our understanding of the epidemiological scenario using short sequence repeats (SSR) to assess the genetic diversity within Paracoccidioides. In silico data mining of 31 reference genomes (27 P. brasiliensis complex and 4 P. lutzii) retrieved from GenBank was performed using GMATA software, resulting in a total of 1,397,207 to 1,470,123 SSR loci identified, displaying a density of 49,480.39 to 50,921.29 SSR/Mb, with most dinucleotide motifs (AT/TC) found in all species. Afterward, we developed a panel of 9 highly polymorphic SSR markers suitable for genotyping members of the P. brasiliensis complex and P. lutzii. PCR amplification revealed 36 alleles in 18 Paracoccidioides isolates (P. brasiliensis complex n=5, P. americana n=2, P. restrepiensis n=4, P. venezuelensis n=2, and P. lutzii n=5), demonstrating excellent polymorphic information content (PIC=0.674), expected heterozygosity (H=0.714), and discriminating power (D=0.663). These findings highlight the SSR markers' efficacy in uncovering cryptic genetic diversity and the genetic population structure of Paracoccidioides spp. Population genetic analysis revealed two clusters corresponding to the P. brasiliensis complex (population 1) and P. lutzii (population 2). Dendrograms (Dice, UPGMA) showed well-supported branches with global cophenetic values exceeding 96%. Principal component analysis (PCA) further confirmed a robust population structure (71.6%). These data significantly contribute to understanding genetic diversity and population structure within Paracoccidioides, providing crucial insights into its epidemiology, taxonomy, and ecology. This information will be vital in developing disease-control strategies to manage PCM effectively. In conclusion, SSR markers offer a rapid, reproducible, and discriminatory DNA fingerprinting tool for the genetic characterization of Paracoccidioides isolates. Palavras-chave: Paracoccidioides, SSR, genetic diversity, genetic characterization, epidemiology Agência de fomento:FAPESP | |||||