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High mitochondrial diversity in geographically widespread butterflies of Madagascar: a test of the DNA barcoding approach.
Mol Phylogenet Evol. 2009 Mar; 50(3):485-95.MP

Abstract

The standardized use of mitochondrial cytochrome c oxidase subunit I (COI) gene sequences as DNA barcodes has been widely promoted as a high-throughput method for species identification and discovery. Species delimitation has been based on the following criteria: (1) monophyletic association and less frequently (2) a minimum 10x greater divergence between than within species. Divergence estimates, however, can be inflated if sister species pairs are not included and the geographic extent of variation within any given taxon is not sampled comprehensively. This paper addresses both potential biases in DNA divergence estimation by sampling range-wide variation in several morphologically distinct, endemic butterfly species in the genus Heteropsis, some of which are sister taxa. We also explored the extent to which mitochondrial DNA from the barcode region can be used to assess the effects of historical rainforest fragmentation by comparing genetic variation across Heteropsis populations with an unrelated forest-associated taxon Saribia tepahi. Unexpectedly, generalized primers led to the inadvertent amplification of the endosymbiont Wolbachia, undermining the use of universal primers and necessitating the design of genus-specific COI primers alongside a Wolbachia-specific PCR assay. Regardless of the high intra-specific genetic variation observed, most species satisfy DNA barcoding criteria and can be differentiated in the nuclear phylogeny. Nevertheless, two morphologically distinguishable candidate species fail to satisfy the barcoding 10x genetic distance criterion, underlining the difficulties of applying a standard distance threshold to species delimitation. Phylogeographic analysis of COI data suggests that forest fragmentation may have played an important role in the recent evolutionary diversification of these butterflies. Further work on other Malagasy taxa using both mitochondrial and nuclear data will provide better insight into the role of historical habitat fragmentation in species diversification and may potentially contribute to the identification of priority areas for conservation.

Authors+Show Affiliations

Department of Biological Sciences, Computer Resources Center Building, Room 200, University of New Orleans, New Orleans, LA 70148, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

19056502

Citation

Linares, Marjorie C., et al. "High Mitochondrial Diversity in Geographically Widespread Butterflies of Madagascar: a Test of the DNA Barcoding Approach." Molecular Phylogenetics and Evolution, vol. 50, no. 3, 2009, pp. 485-95.
Linares MC, Soto-Calderón ID, Lees DC, et al. High mitochondrial diversity in geographically widespread butterflies of Madagascar: a test of the DNA barcoding approach. Mol Phylogenet Evol. 2009;50(3):485-95.
Linares, M. C., Soto-Calderón, I. D., Lees, D. C., & Anthony, N. M. (2009). High mitochondrial diversity in geographically widespread butterflies of Madagascar: a test of the DNA barcoding approach. Molecular Phylogenetics and Evolution, 50(3), 485-95. https://doi.org/10.1016/j.ympev.2008.11.008
Linares MC, et al. High Mitochondrial Diversity in Geographically Widespread Butterflies of Madagascar: a Test of the DNA Barcoding Approach. Mol Phylogenet Evol. 2009;50(3):485-95. PubMed PMID: 19056502.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - High mitochondrial diversity in geographically widespread butterflies of Madagascar: a test of the DNA barcoding approach. AU - Linares,Marjorie C, AU - Soto-Calderón,Iván D, AU - Lees,David C, AU - Anthony,Nicola M, Y1 - 2008/11/21/ PY - 2008/05/16/received PY - 2008/11/06/revised PY - 2008/11/11/accepted PY - 2008/12/6/pubmed PY - 2009/3/6/medline PY - 2008/12/6/entrez SP - 485 EP - 95 JF - Molecular phylogenetics and evolution JO - Mol Phylogenet Evol VL - 50 IS - 3 N2 - The standardized use of mitochondrial cytochrome c oxidase subunit I (COI) gene sequences as DNA barcodes has been widely promoted as a high-throughput method for species identification and discovery. Species delimitation has been based on the following criteria: (1) monophyletic association and less frequently (2) a minimum 10x greater divergence between than within species. Divergence estimates, however, can be inflated if sister species pairs are not included and the geographic extent of variation within any given taxon is not sampled comprehensively. This paper addresses both potential biases in DNA divergence estimation by sampling range-wide variation in several morphologically distinct, endemic butterfly species in the genus Heteropsis, some of which are sister taxa. We also explored the extent to which mitochondrial DNA from the barcode region can be used to assess the effects of historical rainforest fragmentation by comparing genetic variation across Heteropsis populations with an unrelated forest-associated taxon Saribia tepahi. Unexpectedly, generalized primers led to the inadvertent amplification of the endosymbiont Wolbachia, undermining the use of universal primers and necessitating the design of genus-specific COI primers alongside a Wolbachia-specific PCR assay. Regardless of the high intra-specific genetic variation observed, most species satisfy DNA barcoding criteria and can be differentiated in the nuclear phylogeny. Nevertheless, two morphologically distinguishable candidate species fail to satisfy the barcoding 10x genetic distance criterion, underlining the difficulties of applying a standard distance threshold to species delimitation. Phylogeographic analysis of COI data suggests that forest fragmentation may have played an important role in the recent evolutionary diversification of these butterflies. Further work on other Malagasy taxa using both mitochondrial and nuclear data will provide better insight into the role of historical habitat fragmentation in species diversification and may potentially contribute to the identification of priority areas for conservation. SN - 1095-9513 UR - https://www.unboundmedicine.com/medline/citation/19056502/High_mitochondrial_diversity_in_geographically_widespread_butterflies_of_Madagascar:_a_test_of_the_DNA_barcoding_approach_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1055-7903(08)00543-5 DB - PRIME DP - Unbound Medicine ER -