Abstract
Salamanders found at different localities nearby Mérida city, Venezuela, are thus far reported as Bolitoglossa orestes or B.spongai. However, morphological ambiguities among individuals from several populations of both putative species, besidestheir reported disparate geographical distributions, prompted us to clarify the specific identity of these bolitoglossines throughthe sequence analysis of their corresponding 16S rRNA genes. Seventeen specimens belonging to the vertebrates collection ofUniversidad de Los Andes (CVULA), collected at separated cloud forests in Sierra La Culata (San Eusebio, Macho Capaz andSan Javier del Valle) and Sierra Nevada de Mérida (La Mucuy), were used to extract DNA upon tissue digestion. Sequenceanalysis of the 16S rRNA gene supports a biogeographical scenario where, so far, there is only one salamander species for eachsierra: B. orestes, which is widely distributed in Sierra La Culata, and a so far undescribed species of a Venezuelan bolitogloss-ine apparently restricted to Sierra Nevada de Mérida. Based on our molecular results and an examination of morphological evidence, B. spongai should be considered a synonym of B. orestes.References
Adams, D.C., Berns, C.M., Hozak, K.H. & Wiens, J.J. (2009) Are rates of species diversification correlated with rates of morphological evolution? Proceedings of the Royal Society B, 276, 2729–2738.
Ataroff, M. (2001) Venezuela. In: Kappelle, M. & Brown, A. (Eds.), Bosques Nublados del Neotrópico. Editorial IMBIO, Costa Rica, pp. 397–442.
Ataroff, M. & Sarmiento, L. (2004) Las unidades ecológicas de los Andes de Venezuela. In: La Marca, E. & Soriano, P. (Eds.), Reptiles de Los Andes de Venezuela. Fundación Polar, Codepre-ULA, Fundacite-Mérida, Biogeos, Mérida, Venezuela, pp. 9–26.
Barrio-Amorós, C.L. & Fuentes-Ramos, O. (1999) Bolitoglossa spongai a new species of salamander (Caudata: Plethodontidae) of Venezuelan Andes, with comments on the genus in Venezuela. Acta Biologica Venezuelica, 19, 9–19.
Barrio-Amorós, C.L., García, J. & Fuentes-Ramos, O. (2010) Preliminary data on the natural history and intraspecific variation of the endangered salamander Bolitoglossa spongai in the Venezuelan Andes. Salamandra, 2, 108–113.
Blankers, T., Adams, D.C. & Wiens, J.J. (2012) Ecological radiation with limited morphological diversification in salamanders. Journal of Evolutionary Biology, 25, 634–646.
Boulenger, G.A. (1905) Description of new snakes in the collection of the British Museum. The Annals and Magazine of Natural History, including Zoology, Botany and Geology, Seventh Series, 15, 453–456.
Brame Jr., A.H. & Wake, D.B. (1962) A new plethodontid salamander (genus Bolitoglossa) from Venezuela with redescription of the Ecuadorian salamander B. palmata (Werner). Copeia, 1, 170–177.
Cadenas, D.A., Pérez-Sánchez, A.J., Villa, P.M. & de Ascenção, A.A. (2009) Abundancia relativa, uso del hábitat y dieta de Bolitoglossa orestes (Urodela: Plethodontidae) en una selva nublada andina venezolana. Ecotrópicos, 22, 99–109.
Díaz de Pascual, A., García, J., Mora, A., Kiyota, S. & Escalona, M. (2010) Bolitoglossa orestes and Bolitoglossa spongai: life history. Herpetological Review, 41, 186–187.
Dunn, E.R. (1926) The Salamanders of the Family Plethodontidae, Vol. VII Smith College Fiftieth Anniversary Publications. The Southworth Press of Portland, Maine, 456 pp.
Frost, D.R., Grant, T., Faivovich, J., Bain, R.H., Haas, A., Haddad, C.F.B., de Sá, R.O., Channing, A., Wilkinson, M., Donnellan, S.C., Raxworthy, C.J., Campbell, J.A., Blotto, B.L., Moler, P., Drewes, R.C., Nussbaum, R.A., Lynch, J.D., Green, D.M. & Wheeler, W.C. (2006) The amphibian tree of life. Bulletin of the American Museum of Natural History, 297, 1–370.
García-París, M., Good, D., Parra-Olea, G. & Wake, D. (2000) Biodiversity of Costa Rican salamanders: implication of high levels of genetic differentiation and phylogeographic structure for species formation. Proceedings of the National Academy of Sciences, USA, 97, 1640–1647.
Harris, D.M. & Ayala, S.C. (1987) A new Anadia (Sauria: Teiidae) from Colombia and restoration of Anadia Pamplonensis Dunn to species status. Herpetologica, 43, 182–190.
Huang, X. & Madan, A. (1999) CAP3: a DNA sequence assembly program. Genome Research, 9, 868–877.
Larkin, M.A., Blackshields, G., Brown, N.P., Chenna, R., McGettigan, P.A., McWilliam, H., Valentin, F., Wallace, I.M., Wilm, A., Lopez, R., Thompson, J.D., Gibson, T.J. & Higgins, D.G. (2007) Clustal W and Clustal X version 2.0. Bioinformatics, 23, 2947–2948.
Larson, A. & Chippindale, P. (1993) Molecular approaches to the evolutionary biology of plethodontid salamanders. Herpetologica, 49, 204–215.
Mittermeier, R.A., Robles-Gil, P. & Goettsch-Mittermeier, C. (2005) Megadiversity: Earth’s Biologically Wealthiest Nations. Cemex, Monterrey, Mexico, 501 pp.
Palumbi, S.R. (1996) Nucleic Acids II: the Polymerase Chain Reaction. In: Hillis, D.M. Moritz, C. & Mable, B.K. (Eds.), Molecular Systematics. Sinauer & Associates Inc., Sunderland, Massachusetts, pp. 205–247.
Parra-Olea, G., García-París, M. & Wake, D. (2004) Molecular diversification of salamanders of the tropical American genus Bolitoglossa (Caudata:Plethodontidae) and its evolutionary and biogeographical implications. Biological Journal of the Linnean Society, 81, 325–346.
Röhl, E. (1942) Fauna Descriptiva de Venezuela. Tipografía Americana, Caracas, 431 pp.
Sambrook, J. & Russell, D.W. (2001) Molecular Cloning: a Laboratory Manual, Third Edition. Three Volumes Set, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, Massachusetts.
Schargel, W.E., García-Pérez, J.E. & Smith, E.N. (2002) A new species of Bolitoglossa (Caudata: Plethodontidae) from the Cordillera de Mérida, Venezuela. Proceedings of the Biological Society of Washington, 115, 534–542.
Schargel, W.E. & Rivas-Fuenmayor, G. (2003) Two new country records of salamanders of the genus Bolitoglossa from Colombia and Venezuela. Herpetozoa, 16, 94–95.
Schwarzkopf, T., Riha, S.J., Fahey, T.J. & Degloria, S. (2011) Are cloud forest tree structure and environment related in the Venezuelan Andes? Austral Ecology, 36, 280–289.
Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M. & Kumar, S. (2011) MEGA5: Molecular Evolutionary Genetics Analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28, 2731–2739.
Tilley, S. & Bernardo, J. (1993) Life history evolution in plethodontid salamanders. Herpetologica, 49, 154–163.
Trapido, H. (1942) A new salamander from Venezuela. Boletín de la Sociedad Venezolana de Ciencias Naturales, 8, 297–301.
Valbuena-Gómez, J. (2009) Mérida. In: Pantín, G., Reyes, A., Montero, R., Cunill-Grau, P., Márquez, A., Sotillo, A., Aizpúrua, J. & Fontanillas, G. (Eds.), Geo Venezuela, Vol. 6: Geografía de la División Político Territorial del País. Fundación Empresas Polar, Exlibris, Caracas, pp. 484–561.
Vences, M., Thomas, M., Bonett, R.M. & Vieites, D.R. (2005a) Deciphering amphibian diversity through DNA barcoding: chances and challenges. Philosophical Transactions of the Royal Society B, 360, 1859–1868.
Vences, M., Thomas, M., van der Meijden, A., Chiari, Y. & Vieites, D.R. (2005b) Comparative performance of the 16S rRNA gene in DNA barcoding of amphibians. Frontiers in Zoology, 2, 5.
Xia, Y., Gu, H.-F., Peng, R., Chen, Q., Zheng, Y.-C., Murphy, R.W. & Zeng, X.-M. (2012) COI is better than 16S rRNA for DNA barcoding Asiatic salamanders (Amphibia: Caudata: Hynobiidae). Molecular Ecology Resources, 12, 48–56.