Exploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers

Paracoccidioidomycosis (PCM) is a life-threatening systemic fungal infection acquired after inhalation of Paracoccidioides propagules from the environment. The main agents include members of the P. brasiliensis complex (phylogenetically-defined species S1, PS2, PS3, and PS4) and P. lutzii. DNA-seque...

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Autores principales:	T.N. Roberto, J.A. de Carvalho, M.A. Beale, F. Hagen, M.C. Fisher, R.C. Hahn, Z.P. de Camargo, A.M. Rodrigues
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	AFLP AMOVA Endemic mycosis Genetic diversity Mating-type Paracoccidioides Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/3cc372ec1499474cb695d9aa191e50c0
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spelling	oai:doaj.org-article:3cc372ec1499474cb695d9aa191e50c02021-12-02T04:58:59ZExploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers0166-061610.1016/j.simyco.2021.100131https://doaj.org/article/3cc372ec1499474cb695d9aa191e50c02021-09-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S016606162100018Xhttps://doaj.org/toc/0166-0616Paracoccidioidomycosis (PCM) is a life-threatening systemic fungal infection acquired after inhalation of Paracoccidioides propagules from the environment. The main agents include members of the P. brasiliensis complex (phylogenetically-defined species S1, PS2, PS3, and PS4) and P. lutzii. DNA-sequencing of protein-coding loci (e.g., GP43, ARF, and TUB1) is the reference method for recognizing Paracoccidioides species due to a lack of robust phenotypic markers. Thus, developing new molecular markers that are informative and cost-effective is key to providing quality information to explore genetic diversity within Paracoccidioides. We report using new amplified fragment length polymorphism (AFLP) markers and mating-type analysis for genotyping Paracoccidioides species. The bioinformatic analysis generated 144 in silico AFLP profiles, highlighting two discriminatory primer pairs combinations (#1 EcoRI-AC/MseI-CT and #2 EcoRI-AT/MseI-CT). The combinations #1 and #2 were used in vitro to genotype 165 Paracoccidioides isolates recovered from across a vast area of South America. Considering the overall scored AFLP markers in vitro (67–87 fragments), the values of polymorphism information content (PIC = 0.3345–0.3456), marker index (MI = 0.0018), effective multiplex ratio (E = 44.6788–60.3818), resolving power (Rp = 22.3152–34.3152), discriminating power (D = 0.5183–0.5553), expected heterozygosity (H = 0.4247–0.4443), and mean heterozygosity (Havp = 0.00002–0.00004), demonstrated the utility of AFLP markers to speciate Paracoccidioides and to dissect both deep and fine-scale genetic structures. Analysis of molecular variance (AMOVA) revealed that the total genetic variance (65-66 %) was due to variability among P. brasiliensis complex and P. lutzii (PhiPT = 0.651–0.658, P < 0.0001), supporting a highly structured population. Heterothallism was the exclusive mating strategy, and the distributions of MAT1-1 or MAT1-2 idiomorphs were not significantly skewed (1:1 ratio) for P. brasiliensis s. str. (χ2 = 1.025; P = 0.3113), P. venezuelensis (χ2 = 0.692; P = 0.4054), and P. lutzii (χ2 = 0.027; P = 0.8694), supporting random mating within each species. In contrast, skewed distributions were found for P. americana (χ2 = 8.909; P = 0.0028) and P. restrepiensis (χ2 = 4.571; P = 0.0325) with a preponderance of MAT1-1. Geographical distributions confirmed that P. americana, P. restrepiensis, and P. lutzii are more widespread than previously thought. P. brasiliensis s. str. is by far the most widely occurring lineage in Latin America countries, occurring in all regions of Brazil. Our new DNA fingerprint assay proved to be rapid, reproducible, and highly discriminatory, to give insights into the taxonomy, ecology, and epidemiology of Paracoccidioides species, guiding disease-control strategies to mitigate PCM.T.N. RobertoJ.A. de CarvalhoM.A. BealeF. HagenM.C. FisherR.C. HahnZ.P. de CamargoA.M. RodriguesElsevierarticleAFLPAMOVAEndemic mycosisGenetic diversityMating-typeParacoccidioidesBiology (General)QH301-705.5ENStudies in Mycology, Vol 100, Iss , Pp 100131- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	AFLP AMOVA Endemic mycosis Genetic diversity Mating-type Paracoccidioides Biology (General) QH301-705.5
spellingShingle	AFLP AMOVA Endemic mycosis Genetic diversity Mating-type Paracoccidioides Biology (General) QH301-705.5 T.N. Roberto J.A. de Carvalho M.A. Beale F. Hagen M.C. Fisher R.C. Hahn Z.P. de Camargo A.M. Rodrigues Exploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers
description	Paracoccidioidomycosis (PCM) is a life-threatening systemic fungal infection acquired after inhalation of Paracoccidioides propagules from the environment. The main agents include members of the P. brasiliensis complex (phylogenetically-defined species S1, PS2, PS3, and PS4) and P. lutzii. DNA-sequencing of protein-coding loci (e.g., GP43, ARF, and TUB1) is the reference method for recognizing Paracoccidioides species due to a lack of robust phenotypic markers. Thus, developing new molecular markers that are informative and cost-effective is key to providing quality information to explore genetic diversity within Paracoccidioides. We report using new amplified fragment length polymorphism (AFLP) markers and mating-type analysis for genotyping Paracoccidioides species. The bioinformatic analysis generated 144 in silico AFLP profiles, highlighting two discriminatory primer pairs combinations (#1 EcoRI-AC/MseI-CT and #2 EcoRI-AT/MseI-CT). The combinations #1 and #2 were used in vitro to genotype 165 Paracoccidioides isolates recovered from across a vast area of South America. Considering the overall scored AFLP markers in vitro (67–87 fragments), the values of polymorphism information content (PIC = 0.3345–0.3456), marker index (MI = 0.0018), effective multiplex ratio (E = 44.6788–60.3818), resolving power (Rp = 22.3152–34.3152), discriminating power (D = 0.5183–0.5553), expected heterozygosity (H = 0.4247–0.4443), and mean heterozygosity (Havp = 0.00002–0.00004), demonstrated the utility of AFLP markers to speciate Paracoccidioides and to dissect both deep and fine-scale genetic structures. Analysis of molecular variance (AMOVA) revealed that the total genetic variance (65-66 %) was due to variability among P. brasiliensis complex and P. lutzii (PhiPT = 0.651–0.658, P < 0.0001), supporting a highly structured population. Heterothallism was the exclusive mating strategy, and the distributions of MAT1-1 or MAT1-2 idiomorphs were not significantly skewed (1:1 ratio) for P. brasiliensis s. str. (χ2 = 1.025; P = 0.3113), P. venezuelensis (χ2 = 0.692; P = 0.4054), and P. lutzii (χ2 = 0.027; P = 0.8694), supporting random mating within each species. In contrast, skewed distributions were found for P. americana (χ2 = 8.909; P = 0.0028) and P. restrepiensis (χ2 = 4.571; P = 0.0325) with a preponderance of MAT1-1. Geographical distributions confirmed that P. americana, P. restrepiensis, and P. lutzii are more widespread than previously thought. P. brasiliensis s. str. is by far the most widely occurring lineage in Latin America countries, occurring in all regions of Brazil. Our new DNA fingerprint assay proved to be rapid, reproducible, and highly discriminatory, to give insights into the taxonomy, ecology, and epidemiology of Paracoccidioides species, guiding disease-control strategies to mitigate PCM.
format	article
author	T.N. Roberto J.A. de Carvalho M.A. Beale F. Hagen M.C. Fisher R.C. Hahn Z.P. de Camargo A.M. Rodrigues
author_facet	T.N. Roberto J.A. de Carvalho M.A. Beale F. Hagen M.C. Fisher R.C. Hahn Z.P. de Camargo A.M. Rodrigues
author_sort	T.N. Roberto
title	Exploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers
title_short	Exploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers
title_full	Exploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers
title_fullStr	Exploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers
title_full_unstemmed	Exploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers
title_sort	exploring genetic diversity, population structure, and phylogeography in paracoccidioides species using aflp markers
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/3cc372ec1499474cb695d9aa191e50c0
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Exploring genetic diversity, population structure, and phylogeography in Paracoccidioides species using AFLP markers

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