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Revision and phylogeny of narrow-mouthed treefrogs (Cophyla) from northern Madagascar: integration of molecular, osteological, and bioacoustic data reveals three new species



We provide a revision of microhylid treefrogs of the genus Cophyla, the type genus of the subfamily Cophylinae. A phylogeny inferred from DNA sequences of multiple mitochondrial and nuclear genes, with representatives of all cophyline genera except Madecassophryne and including representatives of the two most divergent intrageneric lineages within Cophyla, placed Cophyla as sister group of Platypelis and confirmed both genera as reciprocally monophyletic. We describe three new Cophyla species based on osteological, morphological and bioacoustic characters as well as genetic differentiation in one nuclear and several mitochondrial markers. As in the vast majority of cophylines, all species of Cophyla emit long, stereotyped repetitions of a single tonal note, and we here consider one of these notes as a call; call duration thus equals note duration and the intervals between calls are named inter-call intervals. Cophyla maharipeo sp. nov. collected in Joffreville and Forêt d’Ambre Special Reserve (adult SVL 22–27 mm) is characterized by having long calls (1166–1346 ms) with long inter-call intervals (2154–3881 ms). Cophyla noromalalae sp. nov. collected in Montagne d’Ambre National Park (adult SVL 22–29 mm) is characterized by having short calls (662–821 ms) and short inter-call intervals (874–1882 ms). Cophyla puellarum sp. nov., also from Montagne d’Ambre National Park, is larger than the other two species (adult SVL 27.3–33.6 mm) and characterized by the shortest calls (326–390 ms) and long inter-call intervals (1961–3996 ms). Osteological analyses based on micro-CT scans and cleared and stained specimens confirms that the shape of the posterior vomer (centrally divided vs. undivided) may be a useful character to diagnose most species as belonging to either Platypelis and Cophyla, and suggest the absence of clavicles (present in Platypelis) is a derived character of most Cophyla. However, clavicles were present in C. puellarum, the only known Cophyla occurring at relatively high elevations (1250–1300 m a.s.l.) while otherwise in northern Madagascar, forests at higher elevations up to 2700 m a.s.l. are occupied by Platypelis species. Cophyla maharipeo was found at relatively low elevations (630–720 m a.s.l.), similar to the three previously known congeners (C. berara, C. occultans, C. phyllodactyla). Cophyla noromalalae occurs at intermediate elevations (900–1050 m a.s.l.). The molecular phylogeny inferred herein suggests that the ancestor of a clade containing all Cophyla species except C. puellarum evolved a modified shoulder girdle structure without ossified clavicles, and adapted to low-elevation habitats.



Amphibia, Anura, Microhylidae, Platypelis, conservation status, integrative taxonomy, Montagne d'Ambre, micro-CT scanning

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Andreone, F., Cadle, J.E., Cox, N., Glaw, F., Nussbaum, R.A., Raxworthy, C.J., Stuart, S., Vallan, D. & Vences, M. (2005a) Species review of amphibian extinction risks in Madagascar: conclusions from the Global Amphibian Assessment. Conservation Biology, 19, 1790–1802.

Andreone, F., Vences, M., Vieites, D.R., Glaw, F. & Meyer, A. (2005b) Recurrent ecological adaptations revealed through a molecular analysis of the secretive cophyline frogs of Madagascar. Molecular Phylogenetics and Evolution, 34, 315–322.

Andreone, F., Cox, N., Glaw, F., Köhler, J., Rabibisoa, N.H.C., Randriamahazo, H., Randrianasolo, H., Raxworthy, C.J., Stuart, S.N., Vallan, D. & Vences, M. (2008) Update of the Global Amphibian Assessment for Madagascar in light of new species discoveries, nomenclature changes, and new field information. In: Andreone, F. (Ed.), A Conservation Strategy for the Amphibians of Madagascar. Monografie del Museo Regionale di Scienze Naturali di Torino, 45, pp. 419–438.

Blommers-Schlösser, R.M.A. (1975) Observations on the larval development of some Malagasy frogs, with notes on their ecology and biology (Anura: Dyscophinae, Scaphiophryninae and Cophylinae). Beaufortia, 24, 7–26.

Blommers-Schlösser, R.M.A. & Blanc, C.P. (1991) Amphibiens (première partie). Faune de Madagascar, 75, 1–379.

Boettger, O. (1880) Diagnoses Reptilium et Batrachiorum novorum a Carolo Ebenau in insula Nossi-Bé Madagascariensi lectorum. Zoologischer Anzeiger, 3, 279–283.

Bossuyt, F. & Milinkovitch, M.C. (2000) Convergent adaptive radiations in Madagascan and Asian ranid frogs reveal covariation between larval and adult traits. Proceedings of the National Academy of Sciences of the USA, 97, 6585–6590.

Bruford, M.W., Hanotte, O., Brookfield, J.F.Y. & Burke, T. (1992) Single-locus and multilocus DNA fingerprinting. In: Hoelzel, A.R. (Ed.), Molecular genetic analysis of populations: a practical approach. IRL Press, Oxford, pp. 225–270.

Chiari, Y., Vences, M., Vieites, D.R., Rabemananjara, F., Bora, P., Ramilijaona Ravoahangimalala, O. & Meyer, A. (2004) New evidence for parallel evolution of colour patterns in Malagasy poison frogs (Mantella). Molecular Ecology, 13, 3763–3774.

D'Cruze, N., Köhler, J., Franzen, M. & Glaw, F. (2008) A conservation assessment of the amphibians and reptiles of the Foret d'Ambre Special Reserve, north Madagascar. Madagascar Conservation and Development, 3, 44–54.

Dingerkus, G. & Uhler, L.D. (1977) Enzyme clearing of alcian blue stained whole small vertebrates for demonstration of cartilage. Stain Technology, 52, 229–232.

Folmer, O.M., Black, W., Hoeh, Lutz, R. & Vrijenhoek, R. (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biotechnology, 3, 294–299.

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, 1–291.[0001:TATOL]2.0.CO;2

Glaw, F., Köhler, J. & Vences, M. (2012a) A tiny new species of Platypelis from the Marojejy National Park in northeastern Madagascar (Amphibia: Microhylidae). European Journal of Taxonomy, 9, 1–9.

Glaw, F., Köhler, J.T., Townsend, M. & Vences, M. (2012b) Rivaling the World’s smallest reptiles: Discovery of miniaturized and microendemic new species of leaf chameleons (Brookesia) from northern Madagascar. PLoS ONE, 7, e31314.

Glaw, F. & Vences, M. (1992) A Fieldguide to the Amphibians and Reptiles of Madagascar. Vences & Glaw, Köln, 331 pp., 16 colour plates. [Germany]

Glaw, F. & Vences, M. (1994) A Fieldguide to the Amphibians and Reptiles of Madagascar. 2nd Edition. Vences & Glaw, Verlags GbR, Köln, 480 pp. [Germany]

Glaw, F. & Vences, M. (2007) A Field Guide to the Amphibians and Reptiles of Madagascar. 3rd Edition. Vences & Glaw, Verlags GbR, Köln, 496 pp. [Germany]

Guibé, J. (1978) Les batraciens de Madagascar. Bonner zoologische Monographien, 11, 1–140.

Hrbek, T. & Larson, A. (1999) The evolution of diapause in the killifish family Rivulidae (Atherinomorpha, Cyprinodontiformes): a molecular phylogenetic and biogeographic perspective. Evolution, 53, 12001216.

IUCN (2001) IUCN red list categories and criteria. Version 3.1. IUCN Species Survival Commission, IUCN, Gland, Switzerland and Cambridge, United Kingdom, 30 pp.

Kocher, T.D., Thomas, W.K., Meyer, A., Edwards, S.V., Pääbo, S., Villablanca, F.X. & Wilson, A.C. (1989) Dynamics of mitochondrial DNA evolution in animals: Amplification and sequencing with conserved primers. Proceedings of the National Academy of Sciences of the USA, 86, 6196–6200.

Lanfear, R., Calcott, B., Ho, S.Y.W. & Guindon, S. (2012) PartitionFinder: Combined selection of partitioning schemes and substitution models for phylogenetic analyses. Molecular Biology and Evolution, 29, 1695–1701.

Martin, A.P. (1999) Substitution rates of organelle and nuclear genes in sharks: implicating metabolic rate (again). Molecular Biology and Evolution, 16, 996–1002.

Parker, H.W. (1934) A Monograph of the Frogs of the Family Microhylidae. Trustees of British Museum (Natural History), London, i–viii, 1–208.

Perl, R.G.B., Nagy, Z.T., Sonet, G., Glaw, F., Wollenberg, K.C. & Vences, M. (2014) DNA barcoding Madagascar's amphibian fauna. Amphibia-Reptilia, 35, 197–206.

Pyron, R.A. & Wiens, J.J. (2011) A large-scale phylogeny of Amphibia including over 2800 species, and a revised classification of extant frogs, salamanders, and caecilians. Molecular Phylogenetics and Evolution, 61, 543–583.

Rakotoarison, A., Glaw, F., Vieites, D.R., Raminosoa, N.R. & Vences, M. (2012) Taxonomy and natural history of arboreal microhylidfrogs (Platypelis) from the Tsaratanana massif in northern Madagascar, with description of a new species. Zootaxa, 3563, 1–25.

Rambaut, A. & Drummond, A.J. (2007) Tracer: MCMC Trace Analysis Tool. Institute of Evolutionary Biology, University of Edinburgh, Edinburgh.

Raselimanana, A.P. (2008) Herpétofaune des forêts sèches malgaches. Malagasy Nature, 1, 46–75.

Ronquist, F., Teslenko, M., Van der Mark, P., Ayres, D.L., Darling, A., Höhna, S., Larget, B., Liu, L., Suchard, M.A. & Huelsenbeck, J.P. (2012) MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space. Systematic Biology, 61, 539–542.

Rosa, G.M., Andreone, F., Crottini, A., Hauswaldt, J.S., Noël, J., Rabibisoa, N.H., Randriambahiniarime, M.O., Rebelo, R. & Raxworthy, C.J. (2012) The amphibians of the relict Betampona low-elevation rainforest, eastern Madagascar: an application of the integrative taxonomy approach to biodiversity assessments. Biodiversity & Conservation, 21, 1531–1559.

Shen, X.X., Liang, D. & Zhang, P. (2012) The development of three long universal nuclear protein-coding locus markers and their application to osteichthyan phylogenetics with nested PCR. PLoS ONE 7, e39256.

Swofford, D.L. (2002) PAUP* Phylogenetic Analysis using Parsimony *and other Methods. Version 4. Sinauer Associates, 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, 2731–2739.

van der Meijden, A., Vences, M., Hoegg, S., Boistel, R., Channing, A. & Meyer, A. (2007) Nuclear gene phylogeny of narrow-mouthed toads (Family: Microhylidae) and a discussion of competing hypotheses concerning their biogeographical origins. Molecular Phylogenetics and Evolution, 44, 1017–1030.

Vences, M., Andreone, F. & Glaw, F. (2005) A new microhylid frog of the genus Cophyla from a transitional forest in northwestern Madagascar. African Zoology, 40, 143–149.

Vences, M., Glaw, F., Köhler, J. & Wollenberg, K.C. (2010) Molecular phylogeny, morphology and bioacoustics reveal five additional species of arboreal microhylids of the genus Anodonthyla from Madagascar. Contributions to Zoology, 79, 1–32.

Vences, M., Glaw, F. & Marquez, R. (Eds.) (2006) The Calls of the Frogs of Madagascar. 3 Audio CD's and booklet, 44 pp. Barcelona, Alosa.

Vences, M., Vieites, D.R., Glaw, F., Brinkmann, H., Kosuch, J., Veith, M. & Meyer, A. (2003) Multiple overseas dispersal in amphibians. Proceedings of the Royal Society of London B, 270, 2435–2442

Vieites, D.R., Wollenberg, K.C., Andreone, F., Köhler, J., Glaw, F. & Vences, M. (2009) Vast underestimation of Madagascar's biodiversity evidenced by an integrative amphibian inventory. Proceedings of the National Academy of Sciences of the U.S.A, 106, 8267–8272.

Wollenberg, K.C., Vieites, D.R., van der Meijden, A., Glaw, F., Cannatella, D.C. & Vences, M. (2008) Patterns of endemism and species richness in Malagasy cophyline frogs support a key role of mountainous areas for speciation. Evolution, 62, 1890–1907.


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