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Phylogenetic analysis of microsatellite markers further supports the two hybridization events hypothesis as the origin of the Trypanosoma cruzi lineages

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Abstract

To better understand the evolution of the etiologic agent of Chagas disease, we cloned and sequenced 25 alleles from five Tripanosoma cruzi microsatellite markers. The study of the sequences showed highly conserved alleles present in T. cruzi clones belonging to TCI, TCIIc, and TCIIe. This result was also confirmed by the phylogenetic analysis of MCLE01 allele sequences. The examination by capillary electrophoresis of six microsatellite markers from 19 T. cruzi clones showed a high proportion of the alleles found both in the TCI and TCII sublineages. The phylogenetic reconstruction of these 19 clones produced a tree with two major clusters with bootstrap support of 100% and 95%. The first cluster includes T. cruzi clones belonging to the TCI and TCIIa lineages. The second cluster is composed of TCI, TCIIc, TCIId, and TCIIe T. cruzi clones. The analysis of five microsatellite markers in the CLBrener genome showed that almost all the microsatellite markers are synteny; non-Esmeraldo and Esmeraldo haplotypes probably come from the TCIIc and TCIIb lineages. Taken together, our results are in agreement with the two hybridization events hypothesis as the origin of current T. cruzi lineages.

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Acknowledgments

This study was supported by FONDECYT 1070837. We are grateful to Dr. Christian Barnabé from the Institut de Recherche pour le Developpement, Génétique et Evolution des Maladies Infectieuses, UMR 2724 CNRS/IRD, Montpellier, France, for the kind provision of all T. cruzi clones used in this work. We also thank Leonardo Venegas Hermosilla for excellent technical support in the preparation of all the figures.

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Authors and Affiliations

  1. Programa de Biología Celular y Molecular, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Correo, 70086, Santiago-7, Chile

    Juan Venegas, William Coñoepan, Sergio Pichuantes, Sandra Miranda & Gittith Sánchez

  2. Laboratorio de Referencia en Parasitología, Instituto de Salud Pública de Chile, Santiago, Chile

    María Isabel Jercic

  3. Facultad de Odontología, Universidad de Chile, Santiago, Chile

    Marta Gajardo

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  1. Juan Venegas
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  2. William Coñoepan
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  3. Sergio Pichuantes
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  4. Sandra Miranda
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  5. María Isabel Jercic
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  6. Marta Gajardo
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  7. Gittith Sánchez
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Correspondence to Juan Venegas.

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Supplementary data associated with this article can be found in the online version at doi:…Below is the link to the electronic supplementary material.

Fig. S1

Alignments of the SCLE10 microsatellite alleles cloned from the Trypanosoma cruzi clones: CBB cl3 and SC43 cl1. The letters A1–A3 correspond to the different inserts cloned from each T. cruzi clone. CLBrener-Esm (AAHK01001457.1) and CLBrener non-Esmeraldo (AAHK01000386.1) correspond to the respective haplotypes detected in the published data bank of the T. cruzi CLBrener genome. SCLE10-A and SCLE10-B are the sequences of the respective primers used for the cloning of these marker alleles. In the figure is indicated the position of the CA microsatellite repeat. Numbers flanking the sequences are the respective length of each sequence (RTF 57.0 KB)

Fig. S2

Alignments of the SCLE11 microsatellite alleles cloned from the T. cruzi clones: CBB cl3 and SC43 cl1. The letters A1–A3 correspond to the different inserts cloned from each T. cruzi clone. CLBrener-Esm (AAHK01000459.1) and CLBrener non-Esmeraldo (AAHK01002302.1) correspond to the respective haplotypes detected in the published data bank of the T. cruzi CLBrener genome. SCLE11-A and SCLE11B are the sequences of the respective primers used for the cloning of these marker alleles. In the figure is indicated the position of the CA microsatellite repeat. Numbers flanking the sequences are the respective length of each sequence (RTF 57.0 KB)

Fig. S3

Alignments of the MCLG10 microsatellite alleles cloned from the T. cruzi clones: CLBrener and SC43cl1. The letters A1–A2 correspond to the different inserts cloned from each T. cruzi clone. CLBrener-Esm (AAHK01000157.1) and CLBrener non-Esmeraldo (AAHK01000313.1) correspond to the respective haplotypes detected in the published data bank of the T. cruzi CLBrener genome. MCLG10-A and MCLG10-B are the sequences of the respective primers used for the cloning of these marker alleles. In the figure is indicated the position of the CA microsatellite repeat. Numbers flanking the sequences are the respective length of each sequence (RTF 57.0 KB)

Fig. S4

Alignments of the MCL03 microsatellite alleles cloned from the T. cruzi clones: CBBcl3 and SC43cl1. The letters A1–A3 correspond to the different inserts cloned from each T. cruzi clone. CLBrener (AAHK01000726.1) retrieved from the published data bank of the T. cruzi CLBrener genome. MCLG10-A and MCLG10-B are the sequences of the respective primers used for the cloning of these marker alleles. In the figure is indicated the position of the CA microsatellite repeat. Numbers flanking the sequences are the respective length of each sequence (RTF 57.0 KB)

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Venegas, J., Coñoepan, W., Pichuantes, S. et al. Phylogenetic analysis of microsatellite markers further supports the two hybridization events hypothesis as the origin of the Trypanosoma cruzi lineages. Parasitol Res 105, 191–199 (2009). https://doi.org/10.1007/s00436-009-1386-0

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  • DOI: https://doi.org/10.1007/s00436-009-1386-0

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