Abstract
Species boundaries of Microhyla rubra of India and Sri Lanka were assessed using the following criteria: genetic barcoding, morphology, and vocalization. We use a ca. 500 bp fragment of the 16S rRNA mitochondrial gene and show that there is an uncorrected pairwise distance of 2.7−3.2% between the Indian and Sri Lankan populations of M. rubra. We show that they are different in several call characteristics such as, dominant frequency, call duration, call rise time and pulse rate. Morphologically, the Sri Lankan population can be distinguished from the typical M. rubra described from southern India, by a combination of characters: body size, skin texture, and feet dimensions. We recognize the population from Sri Lanka as a new species, Microhyla mihintalei sp. nov., a widely distributed lowland species with an elevational distribution of up to 500 m a.s.l.
References
Biju, S.D., Garg, S., Gururaja, K.V., Shouche, Y. & Walujkar, S.A. (2014a) DNA barcoding reveals unprecedented diversity in Dancing Frogs of India (Micrixalidae, Micrixalus): a taxonomic revision with description of 14 new species. Ceylon Journal of Science (Biological Sciences), 43 (1), 37–123.
http://dx.doi.org/10.4038/cjsbs.v43i1.6850Biju, S.D., Garg, S., Mahony, S., Wijayathilaka, N., Senevirathne, G. & Meegaskumbura, M. (2014b) DNA barcoding, phylogeny and systematics of Golden-backed frogs (Hylarana, Ranidae) of the Western Ghats-Sri Lanka biodiversity hotspot, with the description of seven new species. Contributions to Zoology, 83 (4), 269–335.
Biju, S.D., Van Bocxlaer, I., Mahony, S., Dinesh, K.P., Radhakrishnan, C., Zachariah, A., Giri, V. & Bossuyt, F. (2011) A taxonomic review of the Night Frog genus Nyctibatrachus Boulenger, 1882 in the Western Ghats, India (Anura: Nyctibatrachidae) with description of twelve new species. Zootaxa, 3029, 1–96.
Bossuyt, F., Meegaskumbura, M., Beenaerts, N., Gower, D.J., Pethiyagoda, R., Roelants, K., Mannaert, A., Wilkinson, M., Bahir, M.M., Manamendra-Arachchi, K., Ng, P.K.L., Schneider, C.J., Oommen, O.V. & Milinkovitch, M.C. (2004) Local endemism within the Western Ghats-Sri Lanka biodiversity hotspot. Science, 306 (5695), 479–481.
http://dx.doi.org/10.1126/science.1100167Bossuyt, F., Meegaskumbura, M., Beenaerts, N., Gower, D.J., Pethiyagoda, R., Roelants, K., Mannaert, A., Wilkinson, M., Bahir, M.M., Manamendra-Arachchi, K., Ng, P.K.L., Schneider, C.J., Oomen, O.V. & Milinkovitch, M.C. (2005) Biodiversity in Sri Lanka and the Western Ghats. Science, 308 (5719), 199.
http://dx.doi.org/10.1126/science.308.5719.199bBowatte, G. & Meegaskumbura, M. (2011) Morphology and ecology of Microhyla rubra (Anura: Microhylidae) tadpoles from Sri Lanka. Amphibian and reptile conservation, 5 (2), 22–32.
De Queiroz, K. (1999) The general lineage concept of species and the defining properties of the species category. In: Wilson, R.A. (Ed.), Species: New Interdisciplinary Essays. MIT Press, Cambridge, Massachusetts, pp. 49–89.
Duméril, A.M.C. & Bibron, G. (1841) Erpétologie Genérale ou Histoire Naturelle Complète des Reptiles. Volume 8, Librarie Enclyclopedique de Roret, Paris. [in French]
Dutta, S.K. (1997) Amphibians of India and Sri Lanka (Checklist and Bibliography). Odyssey Publishing House, Bhubaneswar, Orissa, India, 342 pp.
Dutta, S.K. & Manamendra-Arachchi, K. (1996) The Amphibian Fauna of Sri Lanka. Wildlife Heritage Trust of Sri Lanka, Colombo, 232 pp.
Dutta, S.K. & Ray, P. (2000) Microhyla sholigari, a new species of microhylid frog (Anura: Microhylidae) from Karnataka, India. Hamadryad, 25, 38–44.
Fernando, P. & Siriwardhane, M. (1996) Microhyla karunaratnei (Anura: Microhylidae), a new species of frog endemic to Sri Lanka. Journal of South Asian Natural History, 2, 135–142.
Frank, N. & Ramus., E. (1995) Complete Guide to Scientific and Common Names of Amphibians and Reptiles of the World. Pottsville, Pennsylvania, N.G. Publishing Inc.
Frost, D.R. (2015) Amphibian Species of the World: an Online Reference. Version 6.0. American Museum of Natural History, New York, USA. Available from: http://research.amnh.org/herpetology/amphibia/index.html (accessed 13 January 2015)
Howlader, M.S.A., Nair, A., Gopalan, S.V. & Merilä, J. (2015) A New Species of Microhyla (Anura: Microhylidae) from Nilphamari, Bangladesh. PLoS ONE, 10(3), e0119825.
http://dx.doi:10.1371/journal.pone.0119825Jerdon, T.C. (1854) Catalogue of reptiles inhabiting the Peninsular India. Journal of the Asiatic Society of Bengal, 22, 522–534.
Kirtisinghe, P. (1957) The Amphibia of Ceylon. Published by the author, Colombo, xiii+112 pp.
Kirtisinghe, P. (1958) Some hitherto undescribed anuran tadpoles. Ceylon Journal of Science, 1, 171–176.
Manamendra-Arachchi, K. & Pethiyagoda, R. (2001) Ramanella nagaoi, a new tree-hole frog (Microhylidae) from southern Sri Lanka. Journal of South Asian Natural History, 5 (2), 121–133.
Manamendra-Arachchi, K. & Pethiyagoda, R. (2005) The Sri Lankan shrub-frogs of the genus Philautus Gistel, 1848 (Ranidae: Rhacophorinae), with description of 27 new species. In: Yeo, C.J., Ng, P.K.L. & Pethiyagoda, R. (Eds.), Contributions to biodiversity exploration and research in Sri Lanka. The Raffles Bulletin of Zoology, Supplement 12, 163–303.
Meegaskumbura, M., Bossuyt, F., Pethiyagoda, R., Manamendra-Arachchi, K., Bahir, M., Milinkovitch, M.C. & Schneider, C.J. (2002) Sri Lanka: an amphibian hot spot. Science, 298 (5592), 379.
http://dx.doi.org/10.1126/science.298.5592.379Meegaskumbura, M. & Manamendra-Arachchi, K. (2005) Description of eight new species of shrub frogs (Ranidae: Rhacophorinae: Philautus) from Sri Lanka. In: Yeo, C.J., Ng, P.K.L. & Pethiyagoda, R. (Eds.), Contributions to biodiversity exploration and research in Sri Lanka. The Raffles Bulletin of Zoology, Supplement 12, 305–338.
Meegaskumbura, M. & Manamendra-Arachchi, K. (2011) Two new species of shrub frogs (Rhacophoridae: Pseudophilautus) from Sri Lanka. Zootaxa, 2747, 1–18.
Meegaskumbura, M., Manamendra-Arachchi, K., Bowatte, G. & Meegaskumbura, S. (2012) Rediscovery of Pseudophilautus semiruber, a diminutive shrub frog (Rhacophoridae: Pseudophilautus) from Sri Lanka. Zootaxa, 3229, 58–68.
Meegaskumbura, M., Manamendra-Arachchi, K. & Pethiyagoda, R. (2009) Two new species of shrub frogs (Rhacophoridae: Philautus) from the lowlands of Sri Lanka. Zootaxa, 2122, 51–68.
Meegaskumbura, M., Manamendra-Arachchi, K., Schneider, C.J. & Pethiyagoda, R. (2007) New species amongst Sri Lanka’s extinct shrub frogs (Amphibia: Rhacophoridae: Philautus). Zootaxa, 1397, 1–15.
Meegaskumbura, M., Meegaskumbura, S., Bowatte, G., Manamendra-Arachchi, K., Pethiyagoda, R., Hanken, J. & Schneider, C.J. (2010) Taruga (Anura: Rhacophoridae), a new genus of foam-nesting tree frogs endemic to Sri Lanka. Ceylon Journal of Science (Biological Sciences), 39 (2), 75–94.
http://dx.doi.org/10.4038/cjsbs.v39i2.2995Meegaskumbura, M., Senevirathne, G., Wijayathilaka, N., Jayawardena, B., Bandara, C., Manamendra-Arachchi, K. & Pethiyagoda, R. (2015) The Sri Lankan torrent toads (Bufonidae: Adenominae: Adenomus): species boundaries assessed using multiple criteria. Zootaxa, 3911 (2), 245–261.
http://dx.doi.org/10.11646/zootaxa.3911.2.6Palumbi, 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.
Parker, H.W. (1928) The brevicipitid frogs of the genus Microhyla. Annals and Magazine of Natural History, 2, 473–499.
Parker, H.W. (1934) A monograph of the frogs of the family Microhylidae. British Museum (Natural History), London, 208 pp.
Parker, H.W. & Osman-Hill, W.C. (1949) Frogs of the genus Microhyla from Ceylon. Annals and Magazine of Natural History (Series 12), 1 (10), 759–764.
http://dx.doi.org/10.1080/00222934808653944Peloso, P.L., Frost, D.R., Richards, S.J., Rodrigues, M.T., Donnellan, S., Matsui, M., Raxworthy, C.J., Biju, S.D., Lemmon, E.M., Lemmon, A.R. & Wheeler W.C. (2015) The impact of anchored phylogenomics and taxon sampling on phylogenetic inference in narrow mouthed frogs (Anura, Microhylidae). Cladistics, 1–28.
http://dx.doi.org/10.1111/cla.12118Rao, C.R.N. (1918) Notes on tadpoles of Indian Engystomatidae. Records of the Indian Museum, 15, 41–45.
Swofford, D.L. (2002) PAUP*. Phylogenetic analysis using parsimony (*and other methods). Version 4.0b10. Sinauer Associates, Inc., Sunderland, Massachusetts.
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 (10), 2731–2739.
http://dx.doi.org/10.1093/molbev/msr121Vences, M., Glaw, F., Köhler, J. & Wollenberg, K.C. (2010) Molecular phylogeny, morphology and bioacoustics reveal five additional species of arboreal microhylid frogs of the genus Anodonthyla from Madagascar. Contributions to Zoology, 79 (1), 1–32.
Vences, M., Thomas, M., van der Meijden, A., Chiari, Y. & Vieites, D.R. (2005) Comparative performance of the 16S rRNA gene in DNA barcoding of amphibians. Frontiers in Zoology, 2 (1), 5.
http://dx.doi.org/10.1186/1742-9994-2-5Wickramasinghe, L.J.M., Vidanapathirana, D.R., Airyarathne, S., Rajeev, G., Chanaka, A., Pastorini, J., Chathuranga, G. & Wickramasinghe, N. (2013) Lost and found: One of the world's most elusive amphibians, Pseudophilautus stellatus (Kelaart 1853) rediscovered. Zootaxa, 3620 (1), 112–128.
http://dx.doi.org/10.11646/zootaxa.3620.1.5