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Title Description of a New Species of the Genus Leptobrachella Description of a New Species of the Genus Leptobrachella Title (Amphibia, Anura, Megophryidae) from Borneo Author(s) Eto, Koshiro; Matsui, Masafumi; Nishikawa, Kanto Citation Current Herpetology (2015), 34(2): 128-139 Issue Date 2015-08 URL http://hdl.handle.net/2433/202518 © 2015 by The Herpetological Society of Japan; The full-text file will be made open to the public on 1 September 2018 in Right accordance with publisher's 'Terms and Conditions for Self- Archiving'. Type Journal Article Textversion publisher Kyoto University Description of a New Species of the Genus Leptobrachella (Amphibia, Anura, Megophryidae) from Borneo Author(s): Koshiro Eto, Masafumi Matsui and Kanto Nishikawa Source: Current Herpetology, 34(2):128-139. Published By: The Herpetological Society of Japan DOI: http://dx.doi.org/10.5358/hsj.34.128 URL: http://www.bioone.org/doi/full/10.5358/hsj.34.128 BioOne (www.bioone.org) is a nonprofit, online aggregation of core research in the biological, ecological, and environmental sciences. BioOne provides a sustainable online platform for over 170 journals and books published by nonprofit societies, associations, museums, institutions, and presses. Your use of this PDF, the BioOne Web site, and all posted and associated content indicates your acceptance of BioOne’s Terms of Use, available at www.bioone.org/page/terms_of_use. Usage of BioOne content is strictly limited to personal, educational, and non-commercial use. Commercial inquiries or rights and permissions requests should be directed to the individual publisher as copyright holder. BioOne sees sustainable scholarly publishing as an inherently collaborative enterprise connecting authors, nonprofit publishers, academic institutions, research libraries, and research funders in the common goal of maximizing access to critical research. Current Herpetology 34(2): 128–139, August 2015 doi 10.5358/hsj.34.128 © 2015 by The Herpetological Society of Japan Description of a New Species of the Genus Leptobrachella (Amphibia, Anura, Megophryidae) from Borneo Koshiro ETO*, Masafumi MATSUI, and Kanto NISHIKAWA Graduate School of Human and Environmental Studies, Kyoto University, Yoshida Nihonmatsu-cho, Sakyo-ku, Kyoto 606–8501, JAPAN Abstract: We describe a tiny megophryid frog Leptobrachella juliandringi sp. nov. from Northeastern to Central Sarawak, Malaysian Borneo. This species is distributed in hilly and montane regions, and has long been confused with L. mjobergi. Based on the assessment using morphological and bioacoustic approaches, we conclude that the new species is distinct from all the other congeneric species including topotypic L. mjobergi from western Sarawak in possessing a series of elongate glands ventrolaterally on the trunk, broken reticulated marking on the ventrum, strap-like lineae masculinae, and an ­advertisement­ call­ with­ a­ chirp.­ ­ The­ new­ species­ is­ also­ differentiated­ phylogenetically from the other congeners, and tends to form a group with true L. mjobergi but with a large genetic divergence. Key words: New species; MtDNA; Phylogeny; Leptobrachella mjobergi; Borneo; Taxonomy Introduction been conducted since then, but in view of the recent finding of many cryptic amphibian taxa A Southeast Asian megophryid genus in this region including another megophryid Leptobrachella Smith, 1925 is endemic to genus, Leptolalax (Dehling and Matsui, 2013; Borneo and the Natuna Islands, and its Matsui et al., 2014a, b), taxonomic reassess- members are distinguished by their small ment of this genus is strongly required. adult body size, arrow-like projections on Leptobrachella mjobergi is the type species digital tips, and unique larval morphology of the genus (Smith, 1925), and was thought to (Inger, 1966; Malkmus et al., 2002). This genus occur widely in Southwestern to Northern was thought to include three species (L. mjo- Sarawak and West Kalimantan of Borneo bergi Smith, 1925, L. baluensis Smith, 1931, (Dring, 1983; Inger and Stuebing, 1992), although and L. natunae [Günther, 1895]) until Dring some morphological variations are recognized (1983) described L. brevicrus, L. parva, and within the species (Dring, 1983). We exam- L. serasanae, and Inger and Stuebing (1992) ined intraspecific variations in this species added L. palmata. No taxonomic study has from adult morphology, bioacoustics, and mtDNA sequences, and found that some populations form a clade that is heterospecific * Corresponding author. Tel: +81–75–753–2890; with the topotypic L. mjobergi. In this paper Fax: +81–75–753–6846; we describe the clade as a new species. E-mail address: [email protected] ETO ET AL.—A NEW MEGOPHRYID FROG FROM BORNEO 129 Materials and Methods took the following 21 body measurements following Matsui (1984): (1) snout-vent length For molecular analysis, total DNA was (SVL); (2) head length (HL); (3) snout length extracted from tissues preserved in 99% (SL); (4) snout-nostril length (S-NL); (5) ethanol. Methods for DNA extraction, and nostril-eyelid length (N-EL); (6) eye length following amplification and sequencing of (EL, including eyelid); (7) tympanum-eye mtDNA fragments (approximately 450 base length (T-EL); (8) tympanum diameter (TD); pairs [bp] of 16S rRNA) are same as those (9) head width (HW); (10) internarial distance reported by Matsui et al. (2010b). The resul- (IND); (11) interorbital distance (IOD); (12) tant sequences were deposited in GenBank upper eyelid width (UEW); (13) lower arm and (Accession numbers LC056779–056794: Table hand length (LAL) from elbow to tip of third 1). By using the obtained sequences and finger; (14) hand length (HAL); (15) inner published data from GenBank (Table 1), we palmer tubercle length (IMPL); (16) third reconstructed mitochondrial genealogies based finger length (3FL); (17) hindlimb length on the maximum likelihood (ML) method and (HLL); (18) tibia length (TL); (19) foot length Bayesian inference (BI). (FL); (20) inner metatarsal tubercle length For specimens stored in 70% ethanol, we (IMTL); and (21) first toe length (1TOEL), Table 1. Samples used for DNA analysis in this study, and GenBank accession numbers. BORN: BORNEENSIS collection, University Malaysia Sabah; KUHE: Kyoto University, Human and Environmental Studies; NMBE: Naturhistorisches Museum der Burgergemeinde Bern; SRC: Sarawak Research Collections, Forest Research Center Sarawak; SP: Sabah Parks; UNIMAS: Universiti Malaysia Sarawak; ZMH: Zoological Museum of Hamburg. Species Locality Voucher Accession No. Leptobrachella sp. Mulu NP, Sarawak, Borneo, Malaysia SRC 00230/KUHE 49815 LC056779 Leptobrachella sp. Mulu NP, Sarawak, Borneo, Malaysia KUHE 55333 LC056780 Leptobrachella sp. Mulu NP, Sarawak, Borneo, Malaysia KUHE 55337 LC056781 Leptobrachella sp. Mulu NP, Sarawak, Borneo, Malaysia KUHE 55340 LC056782 Leptobrachella sp. Lanjak Entimau, Sarawak, Borneo, Malaysia KUHE 17554 LC056783 Leptobrachella sp. Lanjak Entimau, Sarawak, Borneo, Malaysia KUHE 17557 LC056784 L. “mjobergi” Mulu NP, Sarawak, Borneo, Malaysia ZMH A13129 KJ831306 L. “mjobergi” Mulu NP, Sarawak, Borneo, Malaysia UNIMAS 8889 KJ831307 L. “mjobergi” Mulu NP, Sarawak, Borneo, Malaysia NMBE 1056347 KJ831308 L. mjobergi Gading NP, Sarawak, Borneo, Malaysia KUHE 17064 LC056785 L. mjobergi Gading NP, Sarawak, Borneo, Malaysia KUHE 17207 LC056786 L. mjobergi Gading NP, Sarawak, Borneo, Malaysia KUHE 47872 LC056787 L. mjobergi Gading NP, Sarawak, Borneo, Malaysia KUHE 47876 LC056788 L. mjobergi Gading NP, Sarawak, Borneo, Malaysia KUHE 48879 LC056789 L. parva Mulu NP, Sarawak, Borneo, Malaysia KUHE 55307 LC056790 L. parva Mulu NP, Sarawak, Borneo, Malaysia KUHE 55308 LC056791 L. parva Mulu NP, Sarawak, Borneo, Malaysia UNIMAS 8883 KJ831304 L. parva Mulu NP, Sarawak, Borneo, Malaysia UNIMAS 8623 KJ831305 L. brevicrus Mulu NP, Sarawak, Borneo, Malaysia ZMH A09365 KJ831302 L. brevicrus Mulu NP, Sarawak, Borneo, Malaysia UNIMAS 8957 KJ831303 L. baluensis Tambunan, Sabah, Borneo, Malaysia SP 21604 LC056792 L. baluensis Kampong Lokos, Sabah, Borneo, Malaysia SP 21687 LC056793 L. baluensis Crocker Range NP, Sabah, Borneo, Malaysia BORN 08595 LC056794 L. baluensis Crocker Range NP, Sabah, Borneo, Malaysia ZMH A13128 KJ831301 Leptolalax gracilis Mulu NP, Sarawak, Borneo, Malaysia KUHE 55624 AB847560 Leptobrachium hasseltii Yogyakarta, Java, Indonesia KUHE 42820 AB530424 130 Current Herpetol. 34(2) 2015 ML tree and the summarized BI trees were 3273.866 and 3316.464, respectively. Phylogenetic relationships obtained from both analyses were nearly identical, and only the ML tree is shown in Fig. 2. The species of Leptobrachella analyzed formed a monophyletic group against outgroups (BS=97%/BPP=1.00). Each spe- cies was also shown to be monophyletic, but three sequences labeled as L. mjobergi from Gunung (Gn.=Mt.) Mulu in GenBank were separated from the topotypes of this species Fig. 1. Map of Sarawak, Malaysian Borneo, and embedded in our Leptobrachella sp. from showing the type locality (filled star), the known Sarawak. Overall, samples analyzed tended distribution (filled circles), and the literature record to be split into two groups: One group with (filled triangles) of Leptobrachella juliandringi sp. poor statistical support (BS=61%/BPP=0.62) nov., with the type locality of L. mjobergi (open star). consisted of Leptobrachella sp. and topotypic L. mjobergi; and another group forming a from distal end of inner metatarsal tubercle to clade (BS=91%/BPP=0.99) contained L. tip of first toe. We made all measurements parva, L. brevicrus, and L. baluensis, within with dial calipers to the
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