Most studies on the diversity of amphibians in northeast India were based on classical morpho-taxonomy, which have a high probability of errors in species identification. DNA barcoding of amphibians using the mitochondrial 16S rRNA gene has been used successfully for the identification of species and detection of cryptic species. In the present study, we carry out DNA barcoding of two rhacophorid species of the genus Raorchestes and Kurixalus that are not readily identifiable to the species level based on their morphology. Our analysis on the 16S rRNA gene revealed that these species were Raorchestes cangyuanensis and Kurixalus yangi, species which were recorded for the first time and second time from India respectively. We discuss on the records of R. longchuanensis from Bangladesh and India, which were based on the misidentification of R. cangyuanensis. We therefore propose to delist this species from the faunal list of Bangladesh and India. We further raise a question as to why the population of Kurixalus from Motuo, Xizang province of China was assigned as K. naso and not the population described as K. yangi, which is morphologically similar to K. naso and is also recorded from a locality close to the type locality of K. naso.
Ahmed, Md. F. (2008) Systematics and Ecology of the Rhacophorid Frogs (Anura: Rhacophoridae) of the Northeast India. Unpublished Ph.D Thesis, Utkal University, Bhubaneswar, Orissa, 252 pp.
Farnsworth, E.J., Chu, M., Kress, W.J., Neill, A.K., Best, J.H., Pickering, J., Stevenson, R.D., Courtney, G.W., VanDyk, J.K. & Ellison, A.M. (2013) Next-Generation Field Guides. BioScience, 63 (11), 891–899.
Al-Razi, H., Maria, M., Hasan, S. & Muzaffar, S.B. (2020) First record of Raorchestes longchuanensis Yang and Li, 1978 (Anura: Rhacophoridae) from northeastern Bangladesh suggests wide habitat tolerance. Amphibian & Reptile Conservation, 14 (1), e225, 119–131.
Annandale, N. (1912) Zoological results of the Abor Expedition, 1911–1912. I. Amphibia. Records of the Indian Museum, 8, 7–36.
CEPF [Critical Ecosystem Partnership Fund] (2016) Announcing the World’s 36th Biodiversity Hotspot: The North American Coastal Plain. Critical Ecosystem Partnership Fund. Available from: https://www.cepf.net (accessed 14 May 2020)
Chitale, V.S., Behera, M.D. & Roy, P.S. (2014) Future of Endemic Flora of Biodiversity Hotspots in India. PLoS ONE, 9 (12), e115264.
Das, A., Chetia, M., Dutta, S.K. & Sengupta, S. (2013) A new species of Duttaphrynus (Anura: Bufonidae) from Northeast India. Zootaxa, 3646 (4), 336–348.
Das, A., Garg, S., Hamidy, A., Smith, E.N. & Biju, S.D. (2019) A new species of Micryletta frog (Microhylidae) from Northeast India. PeerJ, 7 (e7012), 1–32.
Dinesh, K.P., Radhakrishnan, C., Channakeshavamurthy, B.H., Deepak, P. & Kulkarni, N.U. (2020) A Checklist of Amphibians of India with IUCN Conservation Status. Zoological Survey of India, Kolkata, 14 pp.
Garg, S., Suyesh, R., Das, S., Bee, M.A. & Biju, S.D. (2021) An integrative approach to infer systematic relationships and define species groups in the shrub frog genus Raorchestes, with description of five new species from the Western Ghats, India. PeerJ, 9 (e10791), 1–78.
Hasan, M., Islam, M.M., Khan, M.M.R., Alam, M.S., Kurabayashi, A., Igawa, T., Kuramoto, M. & Sumida, M. (2012) Cryptic Anuran Biodiversity in Bangladesh Revealed by Mitochondrial 16S rRNA Gene Sequences. Zoological Science, 29 (3), 162–172.
Hedges, S.B. (1994) Molecular evidence for the origin of birds. Proceedings of the National Academy of Sciences of the United States of America, 91, 2621–2624.
Humtsoe, L.N., Bordoloi, S., Ohler, A. & Grosjean, S. (2020) Natural history and reproductive data of Kurixalus yangi (Anura: Rhacophoridae) in Nagaland, India. Russian Journal of Herpetology, 27 (5), 257–27.
IUCN Standards and Petitions Committee (2019) Guidelines for Using the IUCN Red List Categories and Criteria. Version 14. Prepared by the Standards and Petitions Committee Available from: http://www.iucnredlist.org/documents/RedListGuidelines.pdf (accessed 24 June 2020)
Kamei, R.G., Gower, D.J., Wilkinson, M. & Biju, S.D. (2013) Systematics of the caecilian family Chikilidae (Amphibia: Gymnophiona) with the description of three new species of Chikila from northeast India. Zootaxa, 3666 (4), 401–435.
Kiyasetuo & Khare, M.K. (1987) A new record of Rhacophorus bisacculus Taylor, from North Eastern hills of India. Journal of the Assam Science Society, 29 (1), 16–19.
Kumar, S., Stecher, G., Li, M., Knyaz, C. & Tamura, K. (2018) MEGA X: Molecular Evolutionary Genetics Analysis across Computing Platforms. Molecular Biology and Evolution, 35 (6), 1547–1549.
Lalronunga, S. & Lalrinchhana, C. (2017) Morphological and molecular characterization of Theloderma moloch (Anura: Rhacophoridae) from Indo-Burma biodiversity hotspot of northeast India. Science Vision, 17 (3), 148–159.
Li, J.-T., Che, J., Murphy, R.W., Zhao, H., Zhao, E.-M., Rao, D.-Q. & Zhang, Y.-P. (2009) New insights to the molecular phylogenetics and generic assessment in the Rhacophoridae (Amphibia: Anura) based on five nuclear and three mitochondrial genes, with comments on the evolution of reproduction. Molecular Phylogenetics and Evolution, 53 (2), 509–522.
Mahony, S., Sengupta, S., Kamei, R.G. & Biju, S.D. (2011) A new low altitude species of Megophrys Kuhl and van Hasselt (Amphibia: Megophryidae), from Assam, Northeast India. Zootaxa, 3059 (1), 36–46.
Mahony, S., Teeling, E.C. & Biju, S.D. (2013) Three new species of horned frogs, Megophrys (Amphibia: Megophryidae), from northeast India, with a resolution to the identity of Megophrys boettgeri populations reported from the region. Zootaxa, 3722 (2), 143–169.
Mahony, S., Kamei, R.G., Teeling, E.C. & Biju, S.D. (2018) Cryptic diversity within the Megophrys major species group (Amphibia: Megophryidae) of the Asian Horned Frogs: Phylogenetic perspectives and a taxonomic revision of South Asian taxa, with descriptions of four new species. Zootaxa, 4523 (1), 1–96.
Mahony, S., Kamei, R.G., Teeling, E.C. & Biju, S.D. (2020) Taxonomic review of the Asian Horned Frogs (Amphibia: Megophrys Kuhl & Van Hasselt) of Northeast India and Bangladesh previously misidentified as M. parva (Boulenger), with descriptions of three new species. Journal of Natural History, 54 (1–4), 119–194.
Mathew, R. & Sen, N. (2008) Rediscovery of Rhacophorus naso Annandale, 1912 (Amphibia: Anura: Rhacophoridae) from Mizoram, North East India. Records of the Zoological Survey of India, 108, 41–42.
Mathew, R. & Sen, N. (2009) On the occurrence of Philautus namdaphaensis Sarkar and Sanyal and Rhacophorus naso Annandale (Anura: Rhacophoridae) from Meghalaya, north eastern India. Cobra, 2, 14–16.
Matsui, M., Shimada, T., Liu, W., Maryati, M., Khonsue, W. & Orlov, N. (2006) Phylogenetic relationships of Oriental torrent frogs in the genus Amolops and its allies (Amphibia, Anura, Ranidae). Molecular Phylogenetics and Evolution, 38, 659–666.
Mittermeier, R.A., Robles-Gil, P., Hoffmann, M., Pilgrim, J., Brooks, T., Mittermeier, C.G., Lamoreux, J. & Da Fonseca, G.A.B. (2004) Hotspots revisited: earth’s biologically richest and most endangered terrestrial ecoregions. CEMEX, Mexico City, 390 pp.
Pawar, S. & A. Birand (2001) A survey of amphibians, reptiles and birds in northeast India. CERC Technical Report-6. Center for Ecological Research and Conservation, Mysore, 75 pp.
Pawar, S., Koo, M.S., Kelley, C., Ahmed, M.F., Chaudhuri, S. & Sarkar, S. (2007) Conservation assessment and prioritization of areasin Northeast India: Priorities for amphibians and reptiles. Biological Conservation, 136, 346–361.
Rockney, H.J., Ofori-Boateng, C., Porcino, N. & Leaché, A.D. (2015) A comparison of DNA barcoding markers in West African frogs. African Journal of Herpetology, 64 (2), 135–147.
Roelants, K., Gower, D.J., Wilkinson, M., Loader, S.P., Biju, S.D., Guillaume, K., Moriau, L. & Bossuyt, F. (2007) Global patterns of diversification in the history of modern amphibians. Proceedings of the National Academy of Sciences of the United States of America, 104, 887–892.
Roy, J.K., Begum, R.H. & Ahmed, M.F. (2018) Amphibians of the Dibang River Basin, Arunachal Pradesh: an annotated checklist with distribution records. Journal of Threatened Taxa, 10 (15), 12940–12952.
Roy, P.S., Murthy, M.S.R., Roy, A., Kushwaha, S.P.S., Singh, S., Jha, C.S., Behera, M.D., Joshi, P.K., Jagannathan, C., Karnatak, H.C., Saran, S., Reddy, C.S., Kushwaha, D., Dutt, C.B.S., Porwal, M.C., Sudhakar, S., Srivastava, V.K., Padalia, H., Nandy, S. & Gupta, S. (2013) Forest fragmentation in India. Current Science, 105 (6), 774–780.
Shangpliang, P.W., Hooroo, R.N.K. & Dutta, S.K. (2016) Unique breeding activity and oviposition in Annandale’s high-altitude tree frog, Kurixalus naso (Annandale, 1912) in Meghalaya, North East India. Current Science, 118 (3), 467–472.
Silvestro, D. & Michalak, I. (2012) raxmlGUI: a graphical front-end for RAxML. Organism Diversity and Evolution, 12, 335–337.
Stamatakis, A., Ludwig, T. & Meier, H. (2005) RAxML-III: a fast program for maximum likelihood-based inference of large phylogenetic trees. Bioinformatics, 21, 456–463.
Stamatakis, A. (2006) RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics, 22, 2688–2690.
Stuart, S.N., Hoffmann, M., Chanson, J.S., Cox, N.A., Berridge, R.J., Ramani, P. & Young, B.E. (2008) Threatened amphibians of the world. Lynx Editions, IUCN, Conservation International, Barcelona, Gland and Arlington, 758 pp.
Vences, M., Thomas, M., van der Meijden, A., Chiari, Y. & Vieites, D.R. (2005) Comparative performances of the 16S rRNA gene in DNA barcoding of amphibians. Frontiers in Zoology, 2, 5.
Venkataraman, K., Chattopadhyay, A. & Subramanian, K. (2013) Endemic Animals of India (Vertebrates). Zoological Survey of India, Kolkata, 235 pp.
Williams, K.J., Ford, A., Rosauer, D.F., De Silva, N., Mittermeier, R.A., Bruce, C., Larsen, F.W. & Margules, C. (2011) Forests of East Australia: The 35th Biodiversity Hotspot. In: Zachos, F.E. & Habel, J.C. (Eds.), Biodiversity Hotspots: Distribution and Protection of Priority Conservation Areas. Springer-Verlag, Berlin: pp. 295–310.
Wu, Y.-H., Suwannapoom, C., Xu, K., Chen, J.-M., Jin, J.-Q., Chen, H.-M., Murphy, R.W. & Che, J. (2019) A new species of the genus Raorchestes (Anura: Rhacophoridae) from Yunnan Province, China. Zoological Research, 40, 558–563.
Yu, G., Rao, D., Matsui, M. & Yang, J. (2017) Coalescent-based delimitation outperforms distance-based methods for delineating less divergent species: the case of Kurixalus odontotarsus species group. Scientific Reports, 7, 16124.
Yu, G., Hui, H., Rao, D. & Yang, J. (2018) A new species of Kurixalus from western Yunnan, China (Anura, Rhacophoridae). ZooKeys, 770, 211–226.