Skip to main content Skip to main navigation menu Skip to site footer
Type: Article
Published: 2022-07-21
Page range: 222-236
Abstract views: 439
PDF downloaded: 30

Molecular and morphological identification of frog species collected at Rara Lake in Rara National Park, Nepal

Resources Himalaya Foundation, GPO Box 2448, Naya bato, Ring Road, Lalitpur, Nepal 2Division of Physical and Biological Sciences, Coastal Science and Policy, University of California, Santa Cruz, California, USA
Department of Environmental Science, Khwopa College, Dekocha, Bhaktapur, Nepal
Resources Himalaya Foundation, GPO Box 2448, Naya bato, Ring Road, Lalitpur, Nepal
Department of Wildlife Genetics, Center for Molecular Dynamics Nepal, Kathmandu, Nepal
Department of Wildlife Genetics, Center for Molecular Dynamics Nepal, Kathmandu, Nepal
Department of Wildlife Genetics, Center for Molecular Dynamics Nepal, Kathmandu, Nepal
Département Systematique & Evolution, Museum national d’Histoire naturelle, Paris, France
Département Systematique & Evolution, Museum national d’Histoire naturelle, Paris, France
Department of Vertebrate Zoology, National Museum of Natural History, Smithsonian Institution, Washington, DC, USA 7Department of Biological Sciences, The George Washington University, Washington, DC, USA
Amphibia Meristic characters Molecular phylogeny Paa Rara Lake Nepal

Abstract

This study seeks to clarify the taxonomic identity of three adult frogs and two tadpoles from Rara Lake situated in Rara National Park, Mugu district, Western Nepal, using both phenotypic and mitochondrial sequence data (16S rRNA gene). Based on the molecular data, we determined our specimens belonged to two species; Paa cf. ercepeae (Dubois, 1974) and Paa rarica (Dubois, Matsui, and Ohler, 2001), and this assignment is also supported by morphology. Principal component analysis of phenotypic data clustered species into large-bodied (SVL > 55 mm) and small-bodied (SVL < 55 mm) groups. The molecular data suggest that Paa rarica belonged to a group that contains several undescribed lineages and that Paa liebigii (Günther, 1860) hosts extensive cryptic diversity. Our study also documents that Paa species in the Himalayan region are more widely distributed than previously believed.

 

References

  1. Bhuju, U.R., Shakya, P.R., Basnet, T.B. & Shrestha, S. (2007) Nepal Biodiversity Resource Book: Protected Areas, Ramsar Sites, and World Heritage Sites. International Centre for Integrated Mountain Development: Ministry of Environment, Science and Technology, Government of Nepal, Kathmandu, 128 pp. https://doi.org/10.53055/ICIMOD.475
    Che, J., Hu, J., Zhou, W., Murphy, R.W., Papenfuss, T.J., Chen, M., Rao, D., Li, P. & Zhang, Y. (2009) Phylogeny of the Asian spiny frog tribe Paini (Family Dicroglossidae) sensu Dubois. Molecular Phylogenetics and Evolution, 50 (1), 59–73. https://doi.org/10.1016/j.ympev.2008.10.007
    Che, J., Zhou, W.-W., Hu, J.-S., Yan, F., Papenfuss, T.J., Wake, D.B. & Zhang, Y.-P. (2010) Spiny frogs (Paini) illuminate the history of the Himalayan region and Southeast Asia. Proceedings of the National Academy of Sciences, 107 (31), 13765–13770. https://doi.org/10.1073/pnas.1008415107
    Chen, L., Murphy, R.W., Lathrop, A., Ngo, A., Orlov, N.L., Ho, T. & Somorjai, I.L.M. (2005) Taxonomic chaos in Asian ranid frogs: an initial phylogenetic resolution. Herpetological Journal, 15, 231–243.
    Chettri, B. & Acharya, B.K. (2020) Distribution of amphibians along an elevation gradient in the Eastern Himalaya, India. Basic and Applied Ecology, 47, 57–70. https://doi.org/10.1016/j.baae.2020.07.001
    Dubois, A. (1974a) Diagnoses de trois espèces nouvelles d’amphibiens du Népal. Bulletin de la Société Zoologique de France, 98, 495–497
    Dubois, A. (1974b) Liste commentée d’ Amphibiens récoltés au Népal. Bulletin Du Muséum National d’Histoire Naturelle, Paris, 213 (Zoology 143), 341–411.
    Dubois, A. (1975) Un nouveau sous-genre (Paa) et trois nouvelles espèces du genre Rana. Remarques sur la phylogénie des Ranidés (Amphibiens, Anoures). Bulletin Du Muséum National D’Histoire Naturelle, 324 (231), 1093–1115
    Dubois, A. (1976) Les Grenouilles du sous-genre Paa du Népal (famille Ranidae, genre Rana). Cahiers népalais. Documents. Vol. 6. Centre National de la Recherche Scientifique, Paris, vi + 275 pp.
    Dubois, A. (1981) Biogéographie des Amphibiens de l’Himalaya: état actuel des connaissances. In: Paléogéographie et biogéographie de l’Himalaya et du sous-continent indien. Cahiers népalais. Centre National de la Recherche Scientifique, Paris, pp. 63–74.
    Dubois, A. (1992) Notes sur la classification des Ranidae (Amphibiens, Anoures). Bulletin mensuel de la Société linnéenne de Lyon, 61 (10), 305–352. https://doi.org/10.3406/linly.1992.11011
    Dubois, A. (2000) The influence of man on the distribution of amphibians in the Himalayas of Nepal: an example of critical evaluation of biogeographical data. In: Miehe, G. & Zhang, Y. (Ed.), Environmental changes in high Asia, Marburger geogr. Schriften, 135, pp. 326–345.
    Dubois, A. (2004) Book review. Amphibians of Nepal: a few words of caution. Alytes, 21 (3–4), 174–180.
    Dubois, A. & Matsui, M. (1983) A New Species of Frog (Genus Rana, Subgenus Paa) from Western Nepal (Amphibia: Anura). Copeia, 4, 895. https://doi.org/10.2307/1445091
    Dubois, A., Matsui, M. & Ohler, A. (2001) A replacement name for Rana (Paa) rara Dubois & Matsui, 1983 (Amphibia, Anura, Ranidae, Raninae). Alytes, 19, 2–4. https://doi.org/10.2307/1445091
    Dubois, A., Ohler, A. & Pyron, R.A. (2021) New concepts and methods for phylogenetic taxonomy and nomenclature in zoology, exemplified by a new ranked cladonomy of recent amphibians (Lissamphibia). Megataxa, 5 (1), 1–738. https://doi.org/10.11646/megataxa.5.1.1
    Edgar, R.C. (2004) MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Research, 32 (5), 1792–1797. https://doi.org/10.1093/nar/gkh340
    Frost, D.R. (2020) Amphibian Species of the World: An Online Reference. Version 6. American Museum of Natural History, New York, New York. Available from: https://amphibiansoftheworld.amnh.org/index.php (accessed 21 August 2020)
    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. https://doi.org/10.1206/0003-0090(2006)297[0001:TATOL]2.0.CO;2
    Gautam, B., Chalise, M.K., Thapa, K.B. & Bhattarai, S. (2020) Distributional Patterns of Amphibians and Reptiles in Ghandruk, Annapurna Conservation Area, Nepal. IRCF Reptiles and Amphibians Conservation and Natural History, 27 (1), 18–28. https://doi.org/10.17161/randa.v27i1.14440
    Gurung, S., Gurung, A., Sharma, C.M., Jüttner, I., Tripathee, L., Bajracharya, R.M., Raut, N., Sitaula, B.K., Zhang, Y., Kang, S. & Guo, J. (2018) Hydrochemistry of Lake Rara: A high mountain lake in western Nepal. Lakes & Reservoirs, 23 (2), 87–97. https://doi.org/10.1111/lre.12218
    Gvoždík, V., Moravec, J. & Kratochvíl, L. (2008) Geographic morphological variation in parapatric Western Palearctic tree frogs, Hyla arborea and Hyla savignyi: are related species similarly affected by climatic conditions? Biological Journal of the Linnean Society, 95 (3), 539–556. https://doi.org/10.1111/j.1095-8312.2008.01056.x
    Hofmann, S., Baniya, C.B., Litvinchuk, S.N., Miehe, G., Li, J. & Schmidt, J. (2019a) Phylogeny of spiny frogs Nanorana (Anura: Dicroglossidae) supports a Tibetan origin of a Himalayan species group. Ecology and Evolution, 9 (24), 14498–14511. https://doi.org/10.1002/ece3.5909
    Hofmann, S., Jablonski, D., Litvinchuk, S.N., Masroor, R. & Schmidt, J. (2021) Relict groups of spiny frogs indicate Late Paleogene-Early Neogene trans-tibet dispersal of thermophile faunal elements. PeerJ, 9, e11793. https://doi.org/10.7717/peerj.11793
    Hofmann, S., Kuhl, H., Baniya, C. B. & Stöck, M. (2019b) Multi-Tissue Transcriptomes Yield Information on High-Altitude Adaptation and Sex-Determination in Scutiger cf. sikimmensis. Genes, 10 (11), 873. https://doi.org/10.3390/genes10110873
    Howlader, M.S.A., Nair, A. & Merilä, J. (2016) A New Species of Frog (Anura: Dicroglossidae) Discovered from the Mega City of Dhaka. PLoS ONE, 11 (3), e0149597. https://doi.org/10.1371/journal.pone.0149597
    Ivanova, N.V., Zemlak, T.S., Hanner, R.H. & Hebert, P.D.N. (2007) Universal primer cocktails for fish DNA barcoding: BARCODING. Molecular Ecology Notes, 7 (4), 544–548. https://doi.org/10.1111/j.1471-8286.2007.01748.x
    Jiang, J., Dubois, A., Ohler, A., Tillier, A., Chen, X., Xie, F. & Stöck, M. (2005) Phylogenetic Relationships of the Tribe Paini (Amphibia, Anura, Ranidae) Based on Partial Sequences of Mitochondrial 12s and 16s rRNA Genes. Zoological Science, 22 (3), 353–362. https://doi.org/10.2108/zsj.22.353
    Khatiwada, J.R., Shu, G., Subedi, T.R., Wang, B., Canatella, D.C., Xie, F. & Jiang, J. (2019) A New Species of Megophryid Frog of the Genus Scutiger from Kangchenjunga Conservation Area, Eastern Nepal. Asian Herpetological Research, 10 (3), 24. https://doi.org/10.16373/j.cnki.ahr.180076
    Khatiwada, J.R., Shu, G.C., Wang, S.H., Thapa, A., Wang, B. & Jiang, J. (2017) A new species of the genus Microhyla (Anura: Microhylidae) from Eastern Nepal. Zootaxa, 4254 (2), 221–239. https://doi.org/10.11646/zootaxa.4254.2.4
    Khatiwada, J.R., Shu, G., Wang, B., Zhao, T., Xie, F. & Jiang, J. (2020) Description of a New Species of Amolops Cope, 1865 (Amphibia: Ranidae) from Nepal and Nomenclatural Validation of Amolops nepalicus Yang, 1991. Asian Herpetological Research, 11 (2), 71–94. https://doi.org/10.16373/j.cnki.ahr.190052
    Khatiwada, J.R., Wang, B., Ghimire, S., Paudel, S. & Jiang, J. (2015) A New Species of the Genus Tylototriton (Amphibia: Urodela: Salamandridae) from Eastern Himalaya. Asian Herpetological Research, 6 (4), 245–256. https://doi.org/10.16373/j.cnki.ahr.140097
    Khatiwada, J.R., Wang, S., Shu, G., Xie, F. & Jiang, J. (2018) The mitochondrial genome of the Microhyla taraiensis (Anura: Microhylidae) and related phylogenetic analyses. Conservation Genetics Resources, 10 (3), 441–444. https://doi.org/10.1007/s12686-017-0844-8
    Khatiwada, J.R., Wang, B., Zhao, T., Xie, F. & Jiang, J. (2021) An Integrative Taxonomy of Amphibians of Nepal: An Updated Status and Distribution. Asian Herpetological Research, 12 (1), 1–35. https://doi.org/j.cnki.ahr.200050
    Mitchell, T., Alton, L.A., White, C.R. & Franklin, C.E. (2012) Relations between Conspecific Density and Effects of Ultraviolet-B Radiation on Tadpole Size in the Striped Marsh Frog: UVBR and Conspecific-Density Effects on Amphibians. Conservation Biology, 26 (6), 1112–1120. https://doi.org/10.1111/j.1523-1739.2012.01906.x
    Nanhoe, L.M.R. & Ouboter, P.E. (1987) The distribution of reptiles and amphibians in the Annapurna-Dhaulagiri region (Nepal). Zoölogische Verhandelingen, Leiden, 240 (12), 1–105.
    Nguyen, L.T., Schmidt, H.A., Von Haeseler, A. & Minh, B.Q. (2015) IQ-TREE: a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Molecular biology and evolution, 32 (1), 268–274. https://doi.org/10.1093/molbev/msu300
    O’Connell, K.A., Aryal, P.C., Sherchan, A.M., Dhakal, B., Chaudhary, H.K., Ranabhat, R. & Karmacharya, D. (2019) A herpetological survey of the Kathmandu Valley, Nepal, and phylogenetic identification of Megophrys (Xenophrys) zhangi. Journal of Natural History, 53 (23–24), 1421–1437. https://doi.org/10.1080/00222933.2019.1655106
    Ohler, A. & Dubois, A. (2006) Phylogenetic relationships and generic taxonomy of the tribe Paini (Amphibia, Anura, Ranidae, Dicroglossinae), with diagnoses of two new genera. Zoosystema, 28 (3), 769–784.
    Okino, T. & Satoh, Y. (1986) Morphology, physics, chemistry and biology of Lake Rara in west Nepal. Hydrobiologia, 140, 125–133. https://doi.org/10.1007/BF00007565
    Priti, H., Naik, C.R., Seshadri, K.S., Vidisha, M.K., Ravikanth, G. & Gururaja, V. (2016) A New Species of Euphlyctis (Amphibia, Anura, Dicroglossidae) from the West Coastal Plains of India. Asian Herpetological Research, 7 (4), 229–241. https://doi.org/10.16373/j.cnki.ahr.160020
    R Core Team (2019) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna. Available from: https://www.R-project.org/ (accessed 4 July 2022)
    Rai, K.R. (2003) Environmental Impacts, Systematics and Distribution of Herpetofauna from East Nepal. PhD Dissertation, Central Department of Zoology, Tribhuvan University, Kathmandu, 512 pp.
    Rai, K.R. (2011) Amphibians of Kangchenjunga-Singalilla Complex, Nepal (A Field Guide for Identification, Species Documentation and Conservation Action). ECS-NEPAL, Kathmandu, 88 pp.
    Roelants, K., Jiang, J. & Bossuyt, F. (2004) Endemic ranid (Amphibia: Anura) genera in southern mountain ranges of the Indian subcontinent represent ancient frog lineages: evidence from molecular data. Molecular Phylogenetics and Evolution, 31 (2), 730–740. https://doi.org/10.1016/j.ympev.2003.09.011
    Rueden, C.T., Schindelin, J., Hiner, M.C., DeZonia, B.E., Walter, A.E., Arena, E.T. & Eliceiri, K.W. (2017) ImageJ2: ImageJ for the next generation of scientific image data. BMC bioinformatics, 18 (1), 529. https://doi.org/10.1186/s12859-017-1934-z
    Schleich, H. & Kästle, W. (2002) Amphibians and Reptiles of Nepal: Biology, Systematics, Field Guide. 1st Edition. A.R.G. Ganter Verlag, Kommanditgesellsch, Ruggell, 1201 pp.
    Sewell, D., Griffiths, R.A., Beebee, T.J.C. & Wilkinson, J.W. (2013) Survey protocols for the British herpetofauna Version 1.0. Amphibian and Reptile Conservation, DICE (University of Kent), Canterbury and University of Sussex, Brighton, 22 pp.
    Shah, K.B. & Tiwari, S. (2004) Herpetofauna of Nepal: A Conservation Companion. IUCN—International Union for Conservation of Nature, Kathmandu, Nepal, 237 pp.
    Shrestha, T.K. (2001) Herpetology of Nepal: A study of amphibians and reptiles of Trans-Himalayan Region of Nepal, India, Pakistan and Bhutan. Bimla Shrestha, Kathmandu, 280 pp.
    Terashima, A. (1984) Three new species of the cyprinid genus Schizothorax from Lake Rara, northwestern Nepal. Japanese Journal of Ichthyology, 31 (2), 122–135.
    Wang, K., Bhattarai, S., Wu, Y., Che, J. & Siler, C.D. (2020) Resurrection of Amolops nepalicus Yang, 1991 (Amphibia: Anura: Ranidae), with Comments on the Record of A. cf. afghanus in Nepal and China and the Validity of Two Other Junior Synonyms of A. marmoratus (Blyth, 1855). Zootaxa, 4819 (1), 143–158. https://doi.org/10.11646/zootaxa.4819.1.7
    Wickham, H. (2016) ggplot2: Elegant Graphics for Data Analysis. Springer-Verlag New York, New York, 260 pp. https://doi.org/10.1007/978-3-319-24277-4_9
    Yagi, H., Maemoku, H., Okamura, M., Matsuoka, H., Osada, T., Teramura, H., Adhikari, D.P. & Dangol, V. (2009) Rara Lake, its bathymetric feature and origin, Jumla District, western Nepal Himalayas. Nepal Geological Society, 155, 59–62.

  2.