Abstract
Megophrys are a group of morphologically conserved, primarily forest-dependent frogs known to harbour cryptic species diversity. In this study, we examined populations of small-sized Megophrys from mid- and high elevation locations in the Hoang Lien Range, northern Vietnam. On the basis of morphological, molecular and bioacoustic data, individuals of these populations differed from all species of Megophrys known from mainland Southeast Asia north of the Isthmus of Kra and from neighbouring provinces in China. Further, the newly collected specimens formed two distinct species-level groups. We herein describe two new species, Megophrys fansipanensis sp. nov. and Megophrys hoanglienensis sp. nov. Both new species are range restricted and likely to be highly threatened by habitat degradation. These discoveries highlight the importance of the Hoang Lien Range for Vietnam’s amphibian diversity.
References
Benson, D.A., Cavanaugh, M., Clark, K., Karsch-Mizrachi, I., Lipman, D.J., Ostell, J. & Sayers, E.W. (2017) GenBank. Nucleic Acids Research, 45 (Database issue), D37–D42.
Bickford, D., Lohman, D.J., Sodhi, N.S., Ng, P.K., Meier, R., Winker, K., Ingram, K.K. & Das, I. (2007) Cryptic species as a window on diversity and conservation. Trends in Ecology & Evolution, 22 (3), 148–155.
Boulenger, G.A. (1887) Description of a new frog species of the genus Megalophrys. Annali del Museo Civico di Storia Naturale di Genova, 2, 512–513.
Boulenger, G.A. (1889) Description of a new batrachian of the genus Leptobrachium obtained by M.L. Feae in the Karens Mountains, Burma. Annali del Museo Civico di Storia Naturale di Genova, 2, 748–750.
Boulenger, G.A. (1893) Concluding report on the reptiles and batrachians obtained in Burma by Signor L. Fea, dealing with the collection made in Pegu and the Karin Hills in 1887–8. Annali del Museo Civico di Storia Naturale di Genova, 2, 304–347 + Pl. VII–XII.
Boulenger, G.A. (1899) On a collection of reptiles and batrachians made by Mr. J. D. La Touche in N.W. Fokien, China. Proceedings of the Zoological Society of London, 1899, 159–172 + pls. XVI–XIX.
Boulenger, G.A. (1903) Description of three new batrachians from Tonkin. Annals and Magazine of Natural History, 7, 186–188.
Boulenger, G.A. (1908) A revision of the oriental pelobatid batrachians (genus Megalophrys). Proceedings of the Zoological Society of London, 1908, 407–430 + Pl. XXII–XXV + fig. 71.
Bourret, R. (1937) Notes herpétologiques sur L’Indochine Française. XIV. Les batraciens de la collection du Laboratoire des Sciences Naturelles de l’Université. Descriptions de quinze espèces ou variétés nouvelles. Annexe au Bulletin Général de l’Instruction Publique, 1937, 5–56.
Brown, R.M. & Richards, S.J. (2008) Two new frogs of the genus Platymantis (Anura: Ceratobatrachidae) from the Isabel Island group, Solomon Islands. Zootaxa, 1888 (1), 47–68.
Chen, J.M., Zhou, W.W., Poyarkov Jr, N.A., Stuart, B.L., Brown, R.M., Lathrop, A., Wang, Y.Y., Yuan, Z.Y., Jiang, K., Hou, M. & Chen, H.M. (2017) A novel multilocus phylogenetic estimation reveals unrecognized diversity in Asian horned toads, genus Megophrys sensu lato (Anura: Megophryidae). Molecular Phylogenetics and Evolution, 106, 28–43.
Cocroft, R.B. & Ryan, M.J. (1995) Patterns of advertisement call evolution in toads and chorus frogs. Animal Behaviour, 49 (2), 283–303.
Darriba, D., Taboada, G.L., Doallo, R. & Posada, D. (2012) jModelTest 2: more models, new heuristics and parallel computing. Nature Methods, 9, 772.
https://doi.org/10.1038/nmeth.2109Delorme, M., Dubois, A., Grosjean, S. & Ohler, A. (2006) Une nouvelle ergotaxinomie des Megophryidae (Amphibia, Anoures). Alytes, 24 (1–4), 6–21.
Deuti, K., Grosjean, S, Nicolas, V., Vasudevan, K.V. & Ohler. A. (2017). Nomenclatural puzzle in early Xenophrys nomina (Anura, Megophryidae) solved with description of two new species from India (Darjeeling hills and Sikkim). Alytes, 34, 20–48.
Duellman, W.E. (1970) The Hylid Frogs of Middle America. Monograph of the Museum of Natural History, University of Kansas, 1, 1–427.
Fei, L., Hu, S.Q., Ye, C.Y. & Huang, Y.Z. (2009) Fauna Sinica. Amphibia. Volume 2. Anura. Chinese Academy of Science, Science Press, Beijing, 957 pp.
Fei, L., Ye, C.Y. & Huang, Y.Z. (1983) Two new subspecies of Megophrys omeimontis Liu from China (Amphibia; Pelobatidae). Acta Herpetologica Sinica, 2 (2), 49–52.
Fei, L., Ye, C.Y. & Huang, Y.Z. (1990) Keys to Chinese amphibians. Chongqing Branch, Publishing House of Science and Technology Materials, Chongqing. Unknown pagination.
Fei, L., Ye, C.Y. & Jiang, J.P. (2012) Colored Atlas of Chinese Amphibians and their Distributions. Sichuan Publishing House of Science & Technology, Chengdu, 620 pp.
Foster, P. (2001) The potential negative impacts of global climate change on tropical montane cloud forests. Earth-Science Reviews, 55 (1), 73–106.
Frost, D.R., Grant, T., Faivovich, J., Bain, R.H., Haas, A., Haddad, C.F., De Sá, R.O., Channing, A., Wilkinson, M., Donnellan, S.C. & Raxworthy, C.J. (2006) The amphibian tree of life. Bulletin of the American Museum of natural History, 1–291.
Frost, D.R. (2018) Amphibian Species of the World: an online reference. Version 6. Available from: http://research.amnh.org/herpetologyamphibia/index.html (accessed 9 April 2018)
Günther, A. (1864) The reptiles of British India. London: The Ray Society.
Hanken, J. (1999) Why are there so many new amphibian species when amphibians are declining? Trends in Ecology & Evolution, 14, 7–8
Hijmans, R.J., Cameron S.E., Parra J.L., Jones P.G. & Jarvis A. (2005) Very high resolution interpolated climate surfaces for global land areas. International Journal of Climatology, 25 (15), 1965–1978.
Hillis, D.M. & Bull, J.J. (1993) An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Systematic Biology, 42, 182–192.
https://doi.org/10.1093/sysbio/42.2.182Hu, S., Zhao, E. & Liu, C.C. (1973) A survey of amphibians and reptiles in Kweichow province, including a herpetofaunal analysis. Acta Zoologica Sinica, 19 (2), 149–178.
Huang, Y.Z. & Fei, L. (1981) Two new amphibians from Tibet. Acta Zootaxonomica Sinica, 6, 211–215.
Inger, R.F. & Romer, J.D. (1961) A new Pelobatid frog of the genus Megophrys from Hong Kong. Chicago Natural History Museum, 39 (46), 533–538.
IUCN (2012) IUCN Red List Categories and Criteria: Version 3.1. Second edition. IUCN, Gland, Switzerland and Cambridge, UK, iv + 32pp.
IUCN SSC Amphibian Specialist Group. (2015a) Leptolalax botsfordi. The IUCN Red List of Threatened Species 2015: e.T73727195A73727483.
http://dx.doi.org/10.2305/IUCN.UK.2015-4.RLTS.T73727195A73727483.enIUCN SSC Amphibian Specialist Group. (2015b) Oreolalax sterlingae. The IUCN Red List of Threatened Species 2015: Available from: e.T76491633A76491638.
http://dx.doi.org/10.2305/IUCN.UK.2015-4.RLTS.T76491633A76491638.enIUCN (2017) The IUCN Red List of Threatened Species 2017-3. Available from: http://www.iucnredlist.org/technical-documents/red-list-training/iucnspatialresources (accessed 20 February 2018)
Jiang, J.P., Xie, F., Fei, L., Ye, C. & Zheng, M. (2001) Mating calls of six forms of pelobatid in Wawu Mountain national forest park, Sichuan, China (Anura: Pelobatidae). Zoological research/" Dong wu xue yan jiu" bian ji wei yuan hui bian ji, 23 (1), 89–94.
Katoh, K., Misawa, K., Kuma, K.I. & Miyata, T. (2002) MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Research, 30, 3059–3066.
Köhler, J., Vieites, D.R., Bonett, R.M., García, F.H., Glaw, F., Steinke, D. & Vences, M. (2005) New amphibians and global conservation: a boost in species discoveries in a highly endangered vertebrate group. BioScience, 55 (8), 693–696.
Kuhl, H. & Van Hasselt, J.C. (1822) Uittreksels uit breieven van de Heeren Kuhl en van Hasselt, aan de Heeren C. J. Temminck, Th. van Swinderen en W. de Haan. Algemeene Konst-en Letter-Bode, 7, 99–104.
Kumar, S., Stecher, G. & Tamura, K. (2016) MEGA7: Molecular Evolutionary Genetics Analysis version 7.0 for bigger datasets. Molecular Biology and Evolution, 33 (7), 1870–1874.
Le, D.T., Pham, A.V., Nguyen, S.H.L., Ziegler, T. & Nguyen, T.Q. (2006) First records of Megophrys daweimontis Rao and Yang, 1997 and Amolops vitreus (Bain, Stuart and Orlov, 2006) (Anura: Megophryidae, Ranidae) from Vietnam. Asian Herpetological Research, 6 (1), 66–72.
Li, C. & Wang, Y.Z. (2008) Taxonomic review of Megophrys and Xenophrys, and a proposal for Chinese species (Megophryidae, Anura). Acta Zootaxonomica Sinica, 33 (1), 104–106.
Li, Y.L., Jin, M.J., Zhao, J., Liu, Z.Y., Wang, Y.Y., & Pang, H. (2014) Description of two new species of the genus Megophrys (Amphibia: Anura: Megophryidae) from Heishiding Nature Reserve, Fengkai, Guangdong, China, based on molecular and morphological data. Zootaxa, 3795 (4), 449–471.
http://dx.doi.org/10.11646/zootaxa.3795.4.5Liu, C.C. (1950) Amphibians of western China. Chicago Natural History Museum, 2, 400 pp.
Mahony, S. (2011) Two new species of Megophrys Kuhl & van Hasselt (Amphibia: Megophryidae), from western Thailand and southern Cambodia. Zootaxa, 2734, 23–39.
Mahony, S., Foley, N., Biju, S.D. & Teeling, E. (2017) Evolutionary history of the Asian Horned Frogs (Megophryinae): integrative approaches to timetree dating in the absence of a fossil record. Molecular Biology and Evolution, 34 (3), 744–771.
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, 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.
http://dx.doi.org/10.11646/zootaxa.3722.2.2Mahony, S., Kamei, R.G., Teeling, E.C. & Biju, S.D. (in press) Cryptic diversity within 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, in press.
Miller, M., Pfeiffer, W. & Schwartz, T. (2010) Creating the CIPRES Science Gateway for inference of large phylogenetic trees. Gateway Computing Environments Workshop, 2010, 1–8.
Munir, M., A. Hamidy, Farajallah, A. & Smith, E.N. (2018) A new Megophrys Kuhl and Van Hasselt (Amphibia: Megophryidae) from southwestern Sumatra, Indonesia. Zootaxa, 4442 (3), 389–412.
http://dx.doi.org/10.11646/zootaxa.4442.3.3Neang, T., Chhin, S., Meang, M. & Hun, S. (2013) Confirmation of three species of megophryid frogs (Amphibia Megophryidae) from the Cardamom Mountains of Southwest Cambodia, with the rediscovery of a long lost species. Cambodian Journal of Natural History, 2013 (2), 66–72.
Nguyen, L.T., Poyarkov, Jr., N.A., Le, D.T, Vo, B.D, Ninh, H.T., Duong, T., Murphy, R.W. & Sang, N.V. (2018) A new species of Leptolalax (Anura: Megophryidae) from Son Tra Peninsula, central Vietnam. Zootaxa, 4388 (1), 1–21.
http://dx.doi.org/10.11646/zootaxa.4388.1.1Nguyen, S.V., Ho, T.C. & Nguyen, T.Q. (2009) Herpetofauna of Vietnam, Edition Chimaira, Frankfurt am Main, 768 pp.
Nguyen, T.Q., Phung, T.M., Le, M.D., Ziegler, T. & Böhme, W. (2013) First record of the genus Oreolalax (Anura: Megophryidae) from Vietnam with description of a new species. Copeia, 2013 (2), 213–22.
Oberhummer, E., Barten, C., Schweizer, M., Das, I., Haas, A. & Hertwig, S.T. (2014) Description of the tadpoles of three rare species of megophryid frogs (Amphibia: Anura: Megophryidae) from Gunung Mulu, Sarawak, Malaysia. Zootaxa, 3835 (1), 059–079.
http://dx.doi.org/10.11646/zootaxa.3835.1.3Ohler, A. (2003) Revision of the genus Ophryophryne Boulenger, 1903 (Megophryidae) with description of two new species. Alytes (1–2), 21, 23–44.
Ohler, A., Marquis, O., Swan, S. & Grosjean, S.T. (2000) Amphibian biodiversity of Hoang Lien Nature Reserve (Lao Cai Province, northern Vietnam) with description of two new species. Herpetozoa, 13 (1/2), 71–87.
Ohler, A., Swan, S.R. & Daltry, J.C. (2002) A recent survey of the amphibian fauna of the Cardamom Mountains, southwest Cambodia with descriptions of three new species. The Raffles Bulletin of Zoology, 50 (2), 465–481.
Ohler, A., Wollenberg, K.C., Grosjean, S., Hendrix, R., Vences, M., Ziegler, T. & Dubois, A. (2011) Sorting out Lalos: description of new species and additional taxonomic data on megophryid frogs from northern Indochina (genus Leptolalax, Megophryidae, Anura). Zootaxa, 3147, 1–83.
Orlov, N.L., Murphy, R.W., Ananjeva, N.B., Ryabov, S.A. & Ho, C.T. (2002) Herpetofauna of Vietnam, a checklist. Part 1. Amphibia. Russian Journal of Herpetology, 9 (2), 81–104.
Orlov, N.L., Lathrop, A., Murphy, R.W. & Ho, T.C. (2013) Frogs of the family Rhacophoridae (Anura: Amphibia) in the northern Hoang Lien Mountains (Mount Fan Si Pan, Sa Pa District, Lao Cai Province), Vietnam. Russian Journal of Herpetology, 8 (1), 17–44.
Orlov, N.L., Poyarkov Jr, N.A. & Nguyen, T.T. (2015) Taxonomic Notes on Megophrys frogs (Megophryidae: Anura) of Vietnam, with description of a new species. Russian Journal of Herpetology, 22 (3), 206–218.
Poyarkov Jr, N.A., Rowley, J.J., Gogoleva, S.I., Vassilieva, A.B., Galoyan, E.A. & Orlov, N.L. (2015) A new species of Leptolalax (Anura: Megophryidae) from the western Langbian Plateau, southern Vietnam. Zootaxa, 3931 (2), 221–252.
http://dx.doi.org/10.11646/zootaxa.3931.2.3Poyarkov Jr, N.A., Duong, T.V., Orlov, N.A., Gogoleva, S.S., Vassilieva, A.B., Nguyen, L.T., Nguyen, V.D.H., Nguyen, S.G., Che, G. & Mahony, S. (2017) Molecular, morphological and acoustic assessment of the genus Ophryophryne (Anura, Megophryidae) from Langbian Plateau, southern Vietnam, with description of a new species. Zookeys, 62, 49–120.
Rao, D.Q. & Yang, D.T. (1997) The karyotypes of Megophryinae (Pelobatidae) with a discussion on their classification and phylogenetic relationships. Asiatic Herpetological Research, 7, 93–102.
Rowley, J.J.L., & Cao, T.T. (2009) A new species of Leptolalax (Anura: Megophryidae) from central Vietnam. Zootaxa, 2198 (1), 51–60.
Rowley, J.J.L., Brown, R., Kusrini, M., Inger, R., Stuart, B., Wogan, G., Thy, N., Chan-ard, T., Trung, C.T. & Diesmos, A., (2010a) Impending conservation crisis for Southeast Asian amphibians. Biology Letters, 6 (3), 336–338.
Rowley, J.J.L., Hoang, D.H., Le, T.T.D., Dau, V.Q. & Cao, T.T. (2010b) A new species of Leptolalax (Anura: Megophryidae) from Vietnam and further information on Leptolalax tuberosus. Zootaxa, 2660, 33–45.
Rowley, J.J.L., Hoang, H.D., Dau, V.Q., Thi Thuy Le, D. & Tien Cao, T. (2012) A new species of Leptolalax (Anura: Megophryidae) from central Vietnam. Zootaxa, 3321, 56–68.
Rowley, J.J.L., Dau, V.Q. & Nguyen, T.T. (2013) A new species of Leptolalax (Anura: Megophryidae) from the highest mountain in Indochina. Zootaxa, 3737 (4), 415–428.
http://dx.doi.org/10.11646/zootaxa.3737.4.5Rowley, J.J.L., Stuart, B.L., Neang, T., Hoang, H.D., Dau, V.Q., Nguyen, T.T. & Emmett, D.A. (2015) A new species of Leptolalax (Anura: Megophryidae) from Vietnam and Cambodia. Zootaxa, 4039 (3), 401–417.
http://dx.doi.org/10.11646/zootaxa.4039.3.1Rowley, J.J.L., Tran, D.T., Le, D.T., Dau, V.Q., Peloso, P.L., Nguyen, T.Q., Hoang, H.D., Nguyen, T.T. & Ziegler, T. (2016) Five new, microendemic Asian Leaf-litter Frogs (Leptolalax) from the southern Annamite mountains, Vietnam. Zootaxa, 4085 (1), 63–102.
http://dx.doi.org/10.11646/zootaxa.4085.1.3Rowley, J.J.L., Dau, V.Q. & Cao, T. (2017) A new species of Leptolalax (Anura: Megophryidae) from Vietnam. Zootaxa, 4273, 61–79.
Schlegel, H. (1858) Handleiding tot de Beoefening der Dierkunde. Volume 2. Breda: Koninklijke Militaire Akademie, 628 pp
Shen, Y.H. (1994) A new pelobatid toad of the genus Megophrys from China (Anura: Pelobatidae). In Proceeding of the sixtieth Anniversary of the founding of China Zoological Society, 603–606.
Smith, M.A. (1921) New or Little‐known reptiles and batrachians from southern Annam (Indo‐China). Proceedings of the Zoological Society of London, 91, 423–440.
Stamatakis, A. (2006) RAxML-VI-HPC: maximum likelihood-based phylogenetic analyses with thousands of taxa and mixed models. Bioinformatics, 22, 2688–2690.
Stamatakis, A., Hoover, P. & Rougemont, J. (2008) A rapid bootstrap algorithm for the RAxML web servers. Systematic biology, 57, 758–771.
Stejneger, L. (1926) Two new tailless amphibians from western China. Proceedings of the biological Society of Washington, 39, 53–54.
Stuart, B.L., Inger, R.F. & Voris, H.K., (2006a) High level of cryptic species diversity revealed by sympatric lineages of Southeast Asian forest frogs. Biology Letters, 2 (3), 470–474.
Stuart, B.L., Chuaynkern, Y., Chan-ard, T. & Inger, R.F. (2006b) Three new species of frogs and a new tadpole from eastern Thailand. Fieldiana Zoology, 111, 1–19.
Stuart, B.L., Rowley, J.J., Tran, D.T.A., Le, D.T.T. & Hoang, H.D. (2011) The Leptobrachium (Anura: Megophryidae) of the Langbian Plateau, southern Vietnam, with description of a new species. Zootaxa, 2804 (1), 25–40.
Tapley, B., Cutajar, T., Mahony, S., Nguyen, T.C., Dau, V.Q., Nguyen, T.T. Luong, V.H. & Rowley, J.J.L. (2017a) The Vietnamese population of Megophrys kuatunensis (Amphibia: Megophryidae) represents a new species of Asian horned frog from Vietnam and southern China. Zootaxa, 4344 (3), 465–492.
http://dx.doi.org/10.11646/zootaxa.4344.3.3Tapley, B., Rowley, J.J.L, Cutajar, Nguyen, T.C. & Luong, V.H. (2017b) A conservation action plan for the amphibians of the Hoang Lien Mountain range, 41 pp.
Tapley, B., Michaels, C.J., Gumbs, R., Böhm, M., Luedtke, J., Pearce-Kelly, P. & Rowley, J.J.L. (2018a) The disparity between species description and conservation assessment: a case study in taxa with high rates of species discovery. Biological Conservation, 220, 209–214.
Tapley, B., Cutajar, T., Nguyen, L.T., Nguyen, C.T., Harding, L., Portway, C., Luong, H.V, Rowley, J.J.L. (2018b) A new locality and elevation extension for Megophrys rubrimera in Bat Xat Nature Reserve, Lao Cai Province, northern Vietnam. Herpetology Notes, 11, 865–868.
Wang, Y.Y., Zhang, T.D., Zhao, J., Sung, Y.H., Yang, J.H., Pang, H. & Zhang, Z. (2012) Description of a new species of the genus Xenophrys Günther, 1864 (Amphibia: Anura: Megophryidae) from Mount Jinggang, China, based on molecular and morphological data. Zootaxa, 3546 (1), 53–67.
Wang, Y., Zhao, J., Yang, J., Zhou, Z., Chen, G. & Liu, Y. (2014) Morphology, molecular genetics, and bioacoustics support two new sympatric Xenophrys toads (Amphibia: Anura: Megophryidae) in Southeast China. PLoS ONE, 9 (4), e93075.
Wang, Y.F., Liu, B.Q., Lyu, Jiang, L., Jin, W., Xu, J.N. & Wu, C.H., (2017a). A new species of the Horn Toad of the genus Xenophrys from Zhejiang, China (Amphibia: Megophryidae). Chinese Journal of Zoology, 52 (1), 19 –5929.
Wang, J., Liu, Z.Y. Lyu, Z.T. Zeng, Z.C. & Wang, Y. (2017b). A new species of the genus Xenophrys (Amphibia: Anura: Megophryidae) from an offshore island in Guangdong Province, southeastern China. Zootaxa, 4324 (3), 541–556.
http://dx.doi.org/10.11646/zootaxa.4324.3.8Yang, J.H., Wang, J. & Wang, Y.Y. (2018) A new species of the genus Megophrys (Anura: Megophryidae) from Yunnan Province, China. Zootaxa, 4413 (2), 325–338.
http://dx.doi.org/10.11646/zootaxa.4413.2.5Ye, C. & Fei, L. (1995) Taxonomic studies on the small type Megophrys in China including descriptions of the new species (subspecies) (Pelobatidae: genus Megophrys). Acta Herpetol. Sinica, 4–5, 72–81.
Zheng, Y.C., Mo, B.H., Liu, Z.J. & Zeng, X.M. (2004) Phylogenetic relationships of Megophryid Genera (Anura: Megophryidae) based on partial sequences of mitochondrial 16 S rRNA gene. Zoological Research, 25 (3), 205–213.
Zhang, Y., Li, G., Xiao, N., Li, J., Pan, T., Wang, H., Zhang, B. & Zhou, J. (2017) A new species of the genus Xenophrys (Amphibia: Anura: Megophryidae) from Libo County, Guizhou, China. Asian Herpetological Research, 8, 75–85.