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Testing Introgressive Hybridization Hypotheses Using Statistical Network Analysis of Nuclear and Cytoplasmic Haplotypes in the Leaf Beetle Timarcha goettingensis Species Complex

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Abstract

Previous studies of leaf beetles (Chrysomelidae) in the Timarcha goettingensis species complex using mitochondrial (cox2) and nuclear (ITS-2 rRNA) markers revealed two main clades confined to the Iberian Peninsula and the rest of Europe but showing incongruent distributions indicative of gene exchange between both groups. Because of the anastomosing nature of hybridization, which disrupts the cladistic structure of character variation, phylogenetic trees might be inappropriate to represent and study this process. Here we test for evidence of hybridization in the T. goettingensis complex by analyzing the extra homoplasy arising in hybrid genomes from the simultaneous analysis of genetically independent markers. Haplotype networks obtained by Templeton’s statistical parsimony analysis were generated for combined (concatenated) cox2 and ITS-2 sequences from 167 individuals of the T. goettingensis complex. Networks were used to detect runs of homoplasious characters physically clustered along a nucleotide sequence, as evidence for recombination between both gene partitions. A hypergeometric tail probability for the chance occurrence of physically clustered character changes on the connections linking networks of genotypes was applied. The test recognized two instances of statistically significant clustering, indicating the presence of cox2–ITS-2 mosaic genotypes and reticulation of both main T. goettingensis clades, supporting the reticulate origin of samples of T. maritima in southwestern France and T. sinuatocollis/T. monserratensis in the eastern Pyrenees. Although the assessment of reticulation in DNA sequences does not provide direct proof for hybridization, the geographical distribution of mosaic genotypes in the vicinity of “pure” genotypes supports the effect of gene flow between the two divergent lineages. The study demonstrates the utility of statistical parsimony networks for the detection of hybrids in the growing number of phylogeographic studies based on multiple gene markers.

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Acknowledgments

The authors thank all colleagues who have contributed to this work with samples during several years of research on the genus Timarcha. Ignacio Ribera (MNCN, Madrid) provided useful comments on a preliminary version of the manuscript. We are indebted to Rafael Zardoya (MNCN, Madrid), Brent Emerson (UEA, Norwich), David Posada (University of Vigo, Vigo), Martin Kreitman (University of Chicago, Chicago), and an anonymous reviewer for their support of our work and for constructive comments and suggestions that significantly improved this study. This work was funded by a Marie Curie Fellowship of the European Community under research contract number HPMF-CT-2000-00744.

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Authors and Affiliations

  1. Molecular Systematics Laboratory, Department of Entomology, The Natural History Museum, London, SW7 5BD

    J. Gómez-Zurita & A. P. Vogler

  2. Department of Biological Sciences, Imperial College London, Silwood Park Campus, Ascot, Berkshire, SL5 7PY, UK

    J. Gómez-Zurita & A. P. Vogler

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  1. J. Gómez-Zurita
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  2. A. P. Vogler
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Correspondence to J. Gómez-Zurita.

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[Reviewing Editor: Dr. Rafael Zardoya]

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Gómez-Zurita, J., Vogler, A.P. Testing Introgressive Hybridization Hypotheses Using Statistical Network Analysis of Nuclear and Cytoplasmic Haplotypes in the Leaf Beetle Timarcha goettingensis Species Complex. J Mol Evol 62, 421–433 (2006). https://doi.org/10.1007/s00239-004-0329-8

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