Out of Borneo - Biogeography Katak

Biological Journal of the Linnean Society, 2017, 120, 371–395. With 7 figures.
Out of Borneo, again and again: biogeography of the

Stream Toad genus Ansonia Stoliczka (Anura:
Bufonidae) and the discovery of the first limestone
cave-dwelling species
Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

L. LEE GRISMER1*, PERRY L. WOOD JR2, ANCHALEE AOWPHOL3, MICHAEL COTA4,
MARTA S. GRISMER1, MATTHEW L. MURDOCH1, CESAR AGUILAR2 and
JESSE L. GRISMER5
1
Herpetology Laboratory, Department of Biology, La Sierra University, 4500 Riverwalk Parkway,
Riverside, CA, 92515, USA
2
Department of Biology, Brigham Young University, 150 East Bulldog Boulevard, Provo, UT, 84602,
USA
3
Department of Zoology, Faculty of Science, Kasetsart University, Bangkok, 10900, Thailand
4
Natural History Museum, National Science Museum, Thailand, Technopolis, Khlong 5, Khlong
Luang, Pathum Thani, 12120, Thailand
5
Department of Biological Sciences, Auburn University, 331 Funchess Hall, Auburn, AL, 36849, USA
Received 8 April 2016; revised 7 July 2016; accepted for publication 7 July 2016
Subsequent to the Miocene (approximately 35 Mya), Borneo has served as an insular refuge and a source of
colonization for a broad range of species emigrating to others parts of Sundaland. A phylogeny-based historical
biogeographical hypothesis for the Stream Toad genus Ansonia supports multiple instances of an out-of-Borneo
scenario. An ancestral range estimation indicates that in situ speciation of Ansonia on the island of Borneo
during the Late Miocene and Pliocene (approximately 2–13 Mya) eventually resulted in an invasion of the
Philippines, Sumatra, and two independent invasions of the Thai-Malay Peninsula. When collecting material for
the biogeographical analysis, a new species of Ansonia, Ansonia khaochangensis sp. nov. was discovered in a
limestone cave from the Khao Chang karst tower in Phangnga Province, in southern Thailand. Ansonia
khaochangensis sp. nov. can be differentiated from all other species of Ansonia by having a unique
combination of morphological and colour pattern characteristics. Phylogenetic evidence based on the
mitochondrial genes 12S and 16S indicates that it is nested within a clade of other species distributed north of
the Isthmus of Kra. The cave lifestyle of this new species is a unique and a significant departure from
lotic environments common to most other species of Ansonia. The reproductive biology of this species is
unknown. © 2016 The Linnean Society of London, Biological Journal of the Linnean Society, 2017,
2016, 120, 371–395.
00, 000–000.
KEYWORDS: BioGeoBEARS – biogeography – karst ecosystem – Sundaland – systematics – Thailand.
INTRODUCTION the focus of biogeographers studying the history of

speciation and range expansion of a broad array of
The complex and dynamic environmental history of
lineages distributed across the physiographically var-
South-east Asia has generated considerable interest
ied region referred to as Sundaland – area west
among evolutionary biologists. Much of this has been
of Wallace’s Line encompassing the large islands of
Borneo, Java, and Sumatra and the hundreds of
smaller intervening islands, as well as the
*Corresponding author. E-mail: lgrismer@lasierra.edu
[Version of Record, published online 15 September 2016; Thai-Malay Peninsula south of the Isthmus of Kra
http://zoobank.org/urn:lsid:zoobank.org:pub:20A5589C-7E6F- (Woodruff, 2010). By far the most comprehensive
4245-860A-3BBB2A20B6DC] biogeographical study of Sundaic biota (de Bruyn
© 2016 The Linnean Society of London, Biological Journal of the Linnean Society, 2017, 120, 371–395 371
372 L. LEE GRISMER ET AL.
et al., 2014), convincingly demonstrates that Indochina it is nested within the phylogeny of other species of
and Borneo have been the major sources of coloniza- Ansonia from the Thai-Malay Peninsula and is a dis-
tion for species immigrating into all other regions of tinctive, separately evolving lineage. These data are
Southeast Asia. The unique tectonic and climatic his- corroborated by morphological analyses that dis-
tory of Borneo in particular, enabled this Sundaic cretely differentiate this population from all other
landmass to serve not only as an insular refuge since congeners. It is therefore described here as new.
the Miocene, but an area rich in in situ speciation.
Of the 61 plant, invertebrate, and vertebrate lin-
eages analyzed by de Bruyn et al. (2014), three were
MATERIAL AND METHODS
widespread, semiaquatic frogs from the families
Dicroglossidae and Ranidae. We wanted to test their P HYLOGENETIC AND BIOGEOGRAPHICAL ANALYSES
hypotheses of a Bornean origin and colonization in Ingroup samples consisted of 105 individuals repre-

the diverse microhabitat-restricted frog genus Anso- senting 28 nominal species and five undescribed spe-
nia (Bufonidae) that ranges throughout much of cies of Ansonia (Table 1). Primary outgroups used to
Sundaland, covering most of the pre-defined areas in root the tree were Ingerophrynus divergens, Ingero-
the study by de Bruyn et al. (2014). Stream toads of phrynus parvus, Leptophryne borbonica, Rentapia
the genus Ansonia Stoliczka range from eastern
Myanmar and western Thailand, southward through
the Thai-Malay Peninsula to Borneo and Sumatra,
and then eastward to the Philippines (Matsui et al.,
2010; Chan et al., 2014). This group contains 28
highly scansorial nominal species occupying a rela-
tively narrow ecological zone in Southeast Asian
rainforests where they are closely tied to wet micro-
habitats; be it a seep of dripping water emerging
from an embankment or an open-canopy, fast-flowing
river or stream (Davis et al., 2016). In all cases, their
distinctive morphology of having relatively narrow
bodies; wide, flat heads; and thin, elongate limbs
bearing long digits with bulbous to expanded tips
comprise adaptations for climbing and perching on
rocks and vegetation in lotic environments (Grismer,
2011). Therefore, it was quite unexpected that, dur-
ing the course of our fieldwork, we discovered a new
population living on the walls of a limestone cave in
southern Thailand.
In Southeast Asia, limestone ecosystems are habi-
tat islands dominated by tower karst formations sur-
rounded by a sea of lowland rainforest. Despite the
restrictive microhabitat uniformity associated with
these ecosystems, they are proving to be a source of
rich, unrealized biodiversity and the frequency with
which new species of often highly specialized reptiles
(primarily gekkonids) are being described shows no
signs of levelling off (Grismer et al., 2016). In the
present study, we report the first limestone cave-
adapted population of the stream toad Ansonia from
a karst tower on the edge of the city of Phang Nga,
Phang Nga Province, southern Thailand (Fig. 1)
that, to our knowledge, represents the first cave-
adapted anuran from Sundaland. This new popula-
tion represents a remarkable ecological departure
from the typical lotic lifestyle characterizing all other
Ansonia (Berry, 1975; Inger & Stuebing, 2005). A
molecular analysis of this new population based on Figure 1. Distribution of species of the Thai-Malay
the mitochondrial genes 12S and 16S indicates that Peninsula clade of Ansonia.
© 2016 The Linnean Society of London, Biological Journal of the Linnean Society, 2017, 120, 371–395
HISTORICAL BIOGEOGRAPHY OF STREAM TOADS, GENUS ANSONIA 373
Table 1. Specimens used for the molecular phylogenetic analyses (voucher specimen numbers are available in the refer-
enced citations)
GenBank
Taxon Range number Reference
Outgroups
Duttaphrynus melanostictus East Malaysia, Sarawak, Marudi AB331714 Matsui et al. (2007)
Duttaphrynus melanostictus West Malaysia, Penang, Penang Island AB435318 Matsui et al. (2010)
Ghatophryne ornata India FJ882797 van Boxclear et al.
(2009)
Ingerophrynus divergens Borneo AB331715 Matsui et al. (2007)

Ingerophrynus parvus West Malaysia, Penang, Penang Island AB746455 Matsui et al. (2007)
Rentapia hosii East Malaysia, Sabah, Tawah AB331717 Matsui et al. (2007)
Rentapia hosii East Malaysia, Sabah, Tawah AB331719 Matsui et al. (2007)
Pelophryne brevipes West Malaysia AB331720 Matsui et al. (2007)
Pelophryne misera East Malaysia, Sabah, Kinabalu AB331721 Matsui et al. (2007)
Pelophryne signata East Malaysia, Sarawak, Kuching AB746456 Matsui et al. (2007)
Phrynoidis asper West Malaysia, Penang, Penang Island AB746454 Matsui et al. (2010)
Phrynoidis juxtsaper East Malaysia, Sarawak, Bario AB331713 Matsui et al. (2007)
Leptophryne borbonica East Malaysia, Sabah, Croker, Ulu Kimanis AB331716 Matsui et al. (2007)
Leptophryne borbonica East Malaysia, Sarawak, Penrissen AB746458 Matsui et al. (2007)
Ingroup
Ansonia albomaculata East Malaysia, Sarawak, Lanjak Entimau AB435303 Matsui et al. (2010)
Ansonia endauensis West Malaysia, Johor, Endau-Rompin AB435257 Matsui et al. (2010)
Ansonia fuliginea East Malaysia, Sabah, Kinabalu, Pakka AB435308 Matsui et al. (2010)
Ansonia fuliginea East Malaysia, Sabah, Kinabalu, Pakka AB435309 Matsui et al. (2010)
Ansonia guibei East Malaysia, Sabah, Kinabalu, Mesilau AB435306 Matsui et al. (2010)
Ansonia guibei East Malaysia, Sabah, Kinabalu, Mesilau AB435307 Matsui et al. (2010)
Ansonia hanitschi East Malaysia, Sabah, Kinabalu, Silau AB331710 Matsui et al. (2010)
Ansonia hanitschi Borneo FJ882794 van Boxclear et al.
(2009)
Ansonia inthanon Thailand, Doi Inthanon AB435253 Matsui et al. (2010)
Ansonia jeetsukumarani West Malaysia, Kelantan, Gunung Stong KF906324 Chan et al. (2014)
Ansonia jeetsukumarani West Malaysia, Pahang, Fraser’s Hill KF435265 Chan et al. (2014)
Ansonia jeetsukumarani West Malaysia, Pahang, Fraser’s Hill KF435266 Chan et al. (2014)
Ansonia khaochangensis Thailand, Phang Nga Province, Phang Nga KX032557 ZMKU AM 01145
sp. nov.
sp. nov.
sp. nov.
sp. nov.
sp. nov.
Ansonia kraensis Thailand, Ranong AB435250 Matsui et al. (2010)
Ansonia latidisca East Malaysia, Sarawak, Penrissen AB746459 Matsui et al. (2012)
Table 1. Continued
GenBank
Ansonia latirostra West Malaysia, Pahang, Sungai Lembing AB435260 Matsui et al. (2010)
Ansonia latirostra West Malaysia, Pahang, Sungai Lembing AB435261 Matsui et al. (2010)
Ansonia leptopus East Malaysia, Sabah, Tawah AB435297 Matsui et al. (2010)
Ansonia leptopus East Malaysia, Sarawak, Gunung Gading AB435298 Matsui et al. (2010)
Ansonia leptopus East Malaysia, Sarawak, Kuching AB746457 Matsui et al. (2010)
Ansonia leptopus East Malaysia, Sabah, Croker, Mahua AB331711 Matsui et al. (2007)

Ansonia longidigita East Malaysia, Sabah, Croker, Ulu Kimanis AB435293 Matsui et al. (2010)
Ansonia longidigita East Malaysia, Sabah, Bunduh Tahan AB435294 Matsui et al. (2010)
Ansonia longidigita East Malaysia, Sarawak, Bario AB435295 Matsui et al. (2010)
Ansonia longidigita East Malaysia, Sabah, Sipitang District DQ283341 Frost et al. (2016)
Ansonia longidigita Borneo FJ882795 van Boxclear et al.
(2009)
Ansonia lumut West Malaysia, Terengganu, Gunung Tebu KF906318 Chan et al. (2014)
Ansonia malayana West Malaysia, Perak, Bukit Larut AB331712 Matsui et al. (2010)
Ansonia malayana West Malaysia, Perak, Bukit Larut AB435264 Matsui et al. (2010)
Ansonia mcgregori Philippines, Mindanao AB435316 Matsui et al. (2010)
Ansonia mcgregori Philippines, Mindanao AB435317 Matsui et al. (2010)
Ansonia minuta East Malaysia, Sarawak, Kuching AB435281 Matsui et al. (2010)
Ansonia muelleri Philippines, Mindanao AMU52740 Graybeal (1997)
Ansonia muelleri Philippines, Mindanao AY325992 Darst & Cannetella
(2004)
Ansonia muelleri Philippines, Mindanao AB435309 Matsui et al. (2010)
Ansonia muelleri Philippines, Mindanao, Davao City AB435310 Matsui et al. (2010)
Ansonia muelleri Philippines, Mindanao, Davao City AB435311 Matsui et al. (2010)
Ansonia muelleri Philippines, Mindanao, Davao, Mt. Apo AB435312 Matsui et al. (2010)
Ansonia muelleri Philippines, Mindanao, Davao, Mt. Apo AB435313 Matsui et al. (2010)
Ansonia muelleri Philippines, Mindanao, Mt. Hamigitan AB435314 Matsui et al. (2010)
Ansonia muelleri Philippines, Mindanao, Mt. Hamigitan AB435315 Matsui et al. (2010)
Ansonia penangensis West Malaysia, Penang, Penang Island AB435262 Matsui et al. (2010)
Ansonia penangensis West Malaysia, Penang, Penang Island AB435263 Matsui et al. (2010)
Ansonia platysoma East Malaysia, Sabah, Croker, Mahua AB435267 Matsui et al. (2010)
Ansonia platysoma East Malaysia, Sabah, Croker, Mahua AB435268 Matsui et al. (2010)
Ansonia platysoma East Malaysia, Sabah, Kinabalu, Sayap AB435269 Matsui et al. (2010)
Ansonia platysoma East Malaysia, Sabah, Kinabalu, Bundu Tahan AB435270 Matsui et al. (2010)
Ansonia platysoma East Malaysia, Sabah, Kinabalu, Bundu Tahan AB435271 Matsui et al. (2010)
Ansonia platysoma East Malaysia, Sabah, Kinabalu, Poring AB435272 Matsui et al. (2010)
Ansonia siamensis Thailand, Khaochong AB435255 Matsui et al. (2010)
Ansonia siamensis Thailand, Khaochong AB435256 Matsui et al. (2010)
Ansonia smeagol West Malaysia, Pahang, Genting Highlands KU84554 Davis et al. (2016)
Ansonia sp. 1 East Malaysia, Sarawak, Lanjak Entimau AB435301 Matsui et al. (2010)
Ansonia sp. 1 East Malaysia, Sarawak, Lanjak Entimau AB435302 Matsui et al. (2010)
Table 1. Continued
GenBank
Ansonia sp. 2 East Malaysia, Sabah, Croker, Ulu Kimanis AB435279 Matsui et al. (2010)
Ansonia sp. 2 East Malaysia, Sabah, Croker, Ulu Kimanis AB435280 Matsui et al. (2010)
Ansonia sp. 3 East Malaysia, Sarawak, Bario AB435273 Matsui et al. (2010)
Ansonia sp. 4 Thailand, Phuket Island AB435254 Matsui et al. (2010)

Ansonia sp. 5 Thailand, Kanchanaburi, Pilok AB435249 Matsui et al. (2010)
Ansonia spinulifer East Malaysia, Sabah, Tawah AB4352284 Matsui et al. (2010)
Ansonia spinulifer East Malaysia, Sabah, Maliau AB4352285 Matsui et al. (2010)
Ansonia spinulifer East Malaysia, Sabah, Croker, Ulu Kimanis AB4352286 Matsui et al. (2010)
Ansonia spinulifer East Malaysia, Sarawak, Kuching AB4352289 Matsui et al. (2010)
Ansonia spinulifer East Malaysia, Sarawak, Sadong River AB4352290 Matsui et al. (2010)
Ansonia spinulifer East Malaysia, Sarawak, Sadong River AB4352291 Matsui et al. (2010)
Ansonia spinulifer East Malaysia, Sarawak, Gunung Gading AB4352292 Matsui et al. (2010)
Ansonia spinulifer Borneo FJ882798 van Boxclear et al.
(2009)
Ansonia thinthinae Myanmar, Tanintharyi Division JN664246 Wilkinson et al. (2012)
Ansonia tiomonica West Malaysia, Pahang, Pulau Tioman AB435259 Matsui et al. (2010)
Ansonia tiomonica West Malaysia, Pahang, Pulau Tioman AB435260 Matsui et al. (2010)
Ansonia torrentis East Malaysia, Sarawak, Gunung Mulu AB435296 Matsui et al. (2010)
Ansonia vidua East Malaysia, Sarawak, Gunung Mulu KJ488546 Hertwig et al. (2014)
hosii, Pelophryne brevipes, Pelophryne misera, and deoxynucleotide pairs (1.5 lM), 2.0 lL of 5 9 buffer
Pelophryne signata based in part on the relation- (1.5 lM), 1.0 lL of MgCl 10 9 buffer (1.5 lM),
ships of Chan et al. (2014), Matsui, Yambun & Sudin 0.18 lL of Taq polymerase (5u/lL), and 7.5 of lL
(2007), Matsui et al. (2010), and Wilkinson, Sellas & H2O. All PCR reactions were carried out on an
Vindum (2012). More distant outgroups used were Eppendorf Mastercycler gradient thermocycler fol-
Duttaphrynus melanostictus, Ghatophryne ornata, lowing the thermal profile in Wilkinson et al. (2012):
Phrynoidis asper, and Phrynoidis juxtasper. initial denaturation at 94 °C for 2 min, followed by
Sequence data were generated for five individuals 35 cycles of a secondary denaturation at 94 °C for
from the new Thai population at Phang Nga. 30 s, annealing at 55 °C for 30 s, and elongation at
Sequences for all other individuals from the ingroup 72 °C for 1.5 min, with a final extension at 72 °C for
and outgroup were downloaded from GenBank and 10 min. PCR products were visualized on a 1.0%
all new sequences were deposited in GenBank agarose gel electrophoresis. PCR products that were
(Table 1). the same size as the targeted regions were purified
Genomic DNA was extracted from liver samples using MANU 30 PCR ultrafiltration (Millipore)
and stored in buffer using the animal tissue protocol plates and the products were re-suspended in diH2O.
in the DNeasyTM tissue kit (Qiagen). A 2439-bp frag- Purified PCR products were sequenced using an ABI
ment of the mitochondrial genes 12S, tRNA-VAL, Big-Dye Terminator, version 3.1 Cycle Sequencing
and 16S was amplified using a double-stranded poly- Kit in an ABI GeneAmp PCR 9700 thermal cycler
merase chain reaction (PCR) under the conditions: using internal primers (Chan et al., 2014). All cycle
1.0 lL (approximately 10–33 lg) of genomic DNA, sequencing reactions were purified using Sephadex
1.0 lL of forward external primer (10 lM), 1.0 lL of G-50 Fine (GE Healthcare) and analyzed on an ABI
reverse external primer (10 lM), 1.0 lL of 3730xl DNA Analyzer at the Brigham Young
University Sequencing center. Sequences were edited amphibians using a wide range of calibrations, taxa,
and aligned in GENEIOUS, version 8.1.5 (Drum- and mitochondrial DNA fragments have inferred
mond et al., 2011). Alignment was constructed using model-corrected sequence divergence rates from 0.8%
the MUSCLE, version 3.831 algorithm (Edgar, 2004) to 1.9% total divergence per Myr (Tan & Wake,
implemented in GENEIOUS, version 8.1.5 (Drum- 1995; Macey et al. 1998; Macey et al., 2001; Craw-
mond et al., 2011). ford, 2003a,b; Wang, Crawford & Bermingham, 2008;
The phylogenetic analysis applied two model-based Sanguila et al., 2011). Our run was calibrated to a
methods: maximum likelihood (ML) and Bayesian 1.4% mutation rate sensu Sanguila et al. (2011) for
inference (BI). The ML analysis was performed using Philippine Ansonia using a lognormal prior distribu-
IQ-TREE (Nguyen et al., 2015) with 1000 bootstrap tion and a mean in real space with an SD of 0.05. A
pseudoreplicates using the ultrafast bootstrap maximum clade credibility tree using mean heights
approximation algorithm. The data set was parti- at the nodes was generated using TREEANNOTA-

tioned by gene and the Bayesian information criterion TOR, version 1.8.0 (Rambaut & Drummond, 2013)
(BIC) implemented in IQ-TREE (Nguyen et al., 2015) with a burn-in of 1001 trees. The BEAST log file was
indicated for 12S and 16S that GTR+I+G4 and visualized and checked in TRACER, version 1.6.0
TIM2+I+G4, respectively, were the best-fit models of (Rambaut et al., 2014) to ensure that effective sam-
evolution. For position 2 (valine), the best fit model of ple size (ESS) values were above 200 for all parame-
evolution was TIM2e+G4. The Bayesian analysis was ters. Because ESS values for the ‘prior’ and
carried out using MrBayes, version 3.2.3. on XSEDE ‘posterior’ were below 200 and the remaining param-
(Ronquist et al., 2012) using CIPRES (Cyberinfras- eters were considerably higher than 200 in a prelimi-
tructure for Phylogenetic Research; Miller, Pfeiffer & nary run using a GTR + Gamma substitution model,
Schwartz, 2010) employing default priors. Two inde- a re-run using the less complex HKY model was uti-
pendent runs were performed with four chains for lized because low ESS values may indicate overpa-
each run: three hot and one cold. The simulation, rameterized substitution models (Grummer, Bryson
which ran for 15 000 000 generations, was sampled & Reeder, 2014). Importantly, however, the diver-
every 15 000 generations using Markov chain Monte gence time estimates did not differ significantly
Carlo (MCMC) analysis, and the first 25% of each run between the GTR + Gamma and the HKY models.
was discarded as burn-in. Stationarity was checked in The time-calibrated BEAST tree was pruned to
TRACER, version 1.6 (Rambaut et al., 2014). We con- include one individual per species at each tip, con-
sidered Bayesian posterior probabilities (BPP) of 0.95 verted to Newick format, and then used to estimate
and above and ML ultrafast bootstrap support values the ancestral range at the nodes of the tree using
(ML) of 90 and above as indicating significant nodal the R package BioGeoBEARS (Matzke, 2013). This
support (Huelsenbeck & Ronquist, 2001; Wilcox et al., allows for both probabilistic inference of ancestral
2002; Minh, Nguyen & von Haeseler, 2013). geographical ranges and statistical comparisons of
A time-calibrated Bayesian Inference analysis was range expansion of different models in a likelihood
constructed in BEAUti, version 1.8.0 (Bayesian Evo- framework so that standard statistical model selec-
lutionary Analysis Utility) and implemented in tion procedures such as the Akaike information crite-
BEAST, version 1.8.0 (Bayesian Evolutionary Analy- rion (AIC) can be applied to allow the data to choose
sis Sampling Trees; Drummond et al., 2012), employ- the best fit model. Available models in BioGeo-
ing an uncorrelated lognormal relaxed clock with BEARS include a likelihood version of the parsi-
unlinked substitution and clock models with mono- mony-based Dispersal Vicariance Analysis DIVA
phyly unchecked for taxon sets. MCMC chains were (‘DIVALIKE’) (Ronquist, 1997), the likelihood-based
run using a Speciation: Birth–Death Process for 150 Dispersal-Extinction Cladogenesis (DEC) model of
million generations and logged every 15 000 genera- LAGRANGE (Ree & Smith, 2008), and the Bayesian-
tions. It has been demonstrated that the third codon based BayArea (‘BAYAREALIKE’) (Landis et al.,
position is susceptible to substitution saturation 2013). Additionally, each model also incorporates
(Zamudio, Jones & Ward, 1997; Carranza et al., founder-effect speciation (+J), which was shown to be
2000; Brandley et al., 2011; Grismer et al., 2015) and particularly important when reconstructing biogeo-
could contribute to overestimating node ages. How- graphical scenarios of insular lineages (Matzke,
ever, Grismer et al. (2015) noted that, although third 2014). A presence or absence species geography file
codon position saturation was evident in their study, was constructed utilizing five discrete areas: the
it was not a significant factor in estimating node northern Thai-Malay Peninsula (i.e. that portion of
ages across various codon and gene partition the Thai-Malay Peninsular north of the Isthmus of
schemes and was consistent with similar node age Kra), the southern Thai-Malay Peninsula, the Philip-
estimates using nuclear genes on the same taxa pines, Borneo, and Sumatra. Each species was
(Heinicke et al., 2011). Empirical studies on allowed to occupy a maximum area of two.
M ORPHOLOGICAL ANALYSIS to the Bayesian tree and thus its topology is pre-
Colour notes were taken from the digital images of sented here with both ML ultrafast boostrap values
living and euthanized specimens prior to preserva- and BPP indicated at the nodes that they share in
tion. The characters measured with a Control Com- common (Fig. 2). The phylogeny indicates that the
pany digimatic caliper to the nearest 0.1 mm were: new population from Phang Nga in southern Thai-
snout–vent length (SVL), from tip of snout to vent; land is embedded within a well-supported (1.00/100)
head length (HL), from posterior margin of mandible clade of other species from the Thai-Malay Peninsula
to tip of snout; head width (HW), measured at the and is the sister species to a well-supported (1.00/97)
level of the jaw articulation; snout length (SL), mea- clade composed of the Thai and Burmese species
sured from the anterior margins of the eyes to the Ansonia sp. 4 and 5 from Phuket Island and Kan-
tip of the snout; snout width (SW), distance between chanaburi, respectively; Ansonia inthanon from
western Thailand; Ansonia kraensis from the Thai-

the anterior margins of the eyes; internarial dis-
tance (IND), measured from the medial, inner mar- Malay Peninsula north of the Isthmus of Kra; and
gins of the nostrils; interorbital diameter (IOD), Ansonia thinthinae from southeastern Myanmar
distance between the medial margins of the upper (Figs 1, 2).
eyelids at their closest point; eye diameter (ED), the The time-calibrated BEAST analysis recovered a
length between anterior and posterior margins of topology very similar to that of the ML analysis. The
the eye; tympanum diameter (TD), the length of the topologies differ only with respect to a well-sup-
vertical axis; manus length (ML), the distance from ported Bornean lineage composed of Ansonia spin-
the proximal edge of the outer metacarpal tubercle ulifer, Ansonia sp. 2, Ansonia minuta, Ansonia
to the tip of the third finger; crus length (CL), the hanitschi, Ansonia vidua, and Ansonia sp. 3. In both
distance from the knee inflection to the tarsal inflec- trees, however, some of the nodes within this lineage
tion; and pes length (PL), measured from the proxi- have low support values and, although this consti-
mal edge of the inner metatarsal tubercle to the tip tutes the weakest section of the tree, it does not
of the fourth toe. A toe webbing formula was effect of overall biogeographical scenario. BioGeo-
employed sensu Savage & Heyer (1997). These char- BEARS model comparisons show that the DIVA,
acters were compared with additional discrete char- DEC, and BayArea models with the +J founder-spe-
acter states for all congeners listed in Davis et al. ciation are favoured over the implementation of
(2016). Specimens examined are listed in the appen- these models without +J, in that they had lower AIC
dix of Davis et al. (2016). The type series is depos- values (Table 2). Among them, the DEC +J model
ited in Zoological Museum at Kasetsart University was the best fit to the data and most likely to infer
(ZMKU AM), Bangkok, Thailand. the correct ancestral range at each node (Fig. 4). It
is noteworthy that all the trees generated in all the
analyses recovered the same ancestral range for each
RESULTS respective node on the trees, thus converging on the
same biogeographical scenario. They varied only in
P HYLOGENETIC RELATIONSHIPS the their nodal probability values and all greatly
The Bayesian and ML analyses produced trees with exceeded 50%.
almost identical topologies, differing only in their The morphological analysis supports the molecular
placement of Ansonia tiomanica. The BI analysis analyses by indicating that the Phang Nga popula-
places A. tiomonica as the sister species to Ansonia tion is a discretely diagnosable lineage separable
smeagol, although this relationship is poorly sup- from all other species of Ansonia and, as such, is
ported (0.64). The ML analysis places A. tiomonica described below (ZooBank ID: urn:lsid:zoobank.org:
as the sister species to Ansonia latirostra (Fig. 2). pub:20A5589C-7E6F-4245-860A-3BBB2A20B6DC).
Although this relationship is weakly supported (62),
it has been suggested previously on the basis of mor-
phology (Grandison, 1972) and was recovered in the T AXONOMY
BEAST analysis (Fig. 3). It was weakly supported in Ansonia khaochangensis sp. nov
Matsui et al. (2010) and Chan et al. (2014) but not in Cave-dwelling stream-toad
Davis et al. (2016). Although the present study and Holotype. Adult female (ZMKU AM 01146) col-
those of Chan et al. (2014) and Davis et al. (2016) lected on 1 August 2015 from Khao Chang karst
consider Ansonia malayana and Ansonia jeetsuku- tower, Phang Nga, Takua Pa District, Phang Nga
marani as well-supported sister species they are all Province, Thailand (8° 260 31.8″N; 98° 300 52.1″E,
unresolved as to how these species are related to 91 m in elevation) by L. Lee Grismer, Perry L. Wood
Ansonia lumut and A. smeagol. In general, the ML Jr, Matthew L. Murdoch, Michael Cota, C esar Agui-
tree had far better support across all nodes compared lar, and Marta S. Grismer (Fig. 5).

Figure 2. Maximum likelihood (ML) consensus phylogram for species of the genus Ansonia. Black dots are nodes with
Bayesian posterior probabilities (BPP) of above 0.95 and ML support values above 90; grey dots denote nodes with ML
ultrafast bootstrap values above 90 but BPP below 0.95; and white dots identify nodes with BPP values above 0.95 but
ML ultrafast bootstrap values below 90.
Paratypes. Five paratypes (ZMKU AM 01145, rounded; no dorsolateral row of enlarged tubercles
001147–50) all have the same collection data as the on sides or back; no oblique flap of skin on either
holotype. Their sexes are listed in Table 3. side of vent; abdomen and gular region coarsely
Diagnosis. Ansonia khaochangensis sp. nov. granular; iris black; no light spots on the gular
can be differentiated from all other species of Anso- region or abdomen; no suborbital or postorbital
nia by the combination of the following characters: white markings; no light interscapular spot; no light
maximum SVL of 35.5 mm (34.0–35.3 mm) for crossbars on hind limbs, no light vertebral stripe.
females and 32.0 mm (31.9–32.0 mm) for males; These character states are scored across all species
snout projecting beyond lower jaw; tympanum visi- of Ansonia in Table 4.
ble; vocal sac opening on right; no white or yellow Description of holotype. Adult female, SVL
tubercle at rictus; no tuberculate, interorbital ridges; 34.2 mm; head longer than wide (HL/HW = 1.16);
finger and toe tips rounded, bulbous but not forming snout shorter than wide (SL/SW = 0.91), extending
discs; first finger not reaching disc of second; web- beyond lower jaw, dorsally convex with a midline
bing formula on foot I 1, II ½–2, III 1½–2, IV 2–2, V depression; canthus rostralis distinct; external nares
1; tarsal ridge present; inner and outer metatarsal open laterally just below canthus, almost terminal
tubercles present; submandibular tubercles absent; on snout; distance between nares less than snout
dorsal tubercles present but small, low, and length (IND/SL = 0.45) and snout width (IND/
Figure 3. BEAST chronogram for species of the genus Ansonia. Numbers at the nodes are mean ages in millions of
years. Bars represent the 95% highest posterior probability (HPD).
SW = 0.41); eyes large (ED/SL = 1.07), protruding diameter (TD/ED = 0.58); choanae subcircular, sepa-
beyond jaw line in dorsal view, diameter slightly rated by a distance larger than their diameter;
greater than snout length, less than one-half size of vomerine ridge and teeth absent; tongue narrow, ter-
interorbital distance (ED/IOD = 0.90); tympanum minating in median point, posterior one-half free;
distinct, circular with horizontal axis less than eye upper eyelid, interorbital region, and dorsal part of
snout and canthus sparsely covered with small,
AIC corrected
raised tubercles bearing keratinized tips; interorbital
ridges absent, paravertebral oval areas present; lores
82.08
51.90
80.69
52.17
80.69
52.17
Table 2. Biogeographical model selection with and without founder-event speciation(+j) based on the lowest Akaike information criterion (AIC) values atuberculate; single row of small spinules on outer
margin of upper eyelid; no large tubercle above ric-
tus; supratympanic fold and parotoid glands absent;
submandibular and mental region bearing small
73.08
47.90
70.30
48.17
70.30
48.17
tubercles; gular region coarsely granular to weakly
AIC
tuberculate; back bearing low, small tubercles; large

isolated tubercles in posttympanic region; flanks
bearing widely scattered, large, low, rounded tuber-
0.0000000
0.1142811
0.0000000
0.1157199
0.0000000
0.1081158
cles; abdomen coarsely granular.

Dorsal surfaces of limbs densely tuberculate,
undersides coarsely granular; undersides of pes and
J
manus weakly tuberculate; forelimbs and fingers

long and slender; finger length from shortest to long-
est I, II, IV, III; basal webbing not extending beyond
1.000000e 9 10–12
1.000000e 9 10–12
3.040188e 9 10–1
1.000000 9 10–12
1.000000 9 10–12
proximal subarticular tubercles; fingertips rounded,

2.83563 9 10–2
slightly dilated but not forming discs; subarticular

d, rate of dispersal; e, rate of extinction; j, relative probability of founder-event speciation at cladogenesis.
tubercles distinct; outer palmar tubercle oval, raised;

hind limbs and toes long and slender (CL/
SVL = 0.45), foot longer than tibia (PL/CL = 0.76);
toe length from longest to shortest I, II, III, V, IV;
e
webbing formula: I 1, II ½–2, III 1½–2, IV 2–2, V 1;

tips rounded, dilated but not forming discs; subartic-
1.000000e 9 10–12
1.000000e 9 10–12
ular tubercles weak; inner metatarsal tubercle elon-

4.474416e 9 10–2
2.001000e 9 10–9
1.738085e 9 10–2
3.504546 9 10–2
gate, raised; outer metatarsal tubercle raised, oval,

slightly smaller than inner.
Coloration in life (Fig. 5). Ground colour of dorsal
surfaces of head, body and hind limbs uniform dark-
brown; sides of head and neck dark-brown; ground
d
colour of flanks, forelimbs, hands, and feet dull-

orange to tan; ankle, knee, and elbow joints dull-
orange; small, yellowish tubercles near rictus, on
parameters
Number of
flanks, and fewer yellowish tubercles on limbs and

postfemoral region; and venter beige, immaculate.
Variations (Fig. 5). All paratypes closely resemble
2
3
2
3
2
3
the holotype in all aspects of coloration and morphol-

ogy. The dorsal ground colour of ZMKU AM 01147 is
slightly darker and the hands and feet are light-tan
34.5
20.9
33.1
21.1
40.3
21.6
LnL
as opposed to being dull-orange. Measurements

showing the variation in size are presented in
Table 3.
Comparisons. Morphological data for species com-
Maximum
parisons were obtained from the data matrix of

of areas
number
Davis et al. (2016). Ansonia khaochangensis sp.

nov. is nested within a monophyletic group com-
2
2
2
2
2
2
posed of 14 other species of Ansonia endemic to the

Thai-Malay Peninsula (Fig. 2). It differs from all
these species by lacking light-coloured crossbars on
BAYAREALIKE+J
BAYAREALIKE
the hind limbs and having a black iris as opposed

DIVALIKE+J
to other colours (Table 4). It can be differentiated

DIVALIKE
from all the Thai-Malay species except Ansonia lat-

iffi by males lacking a large white to yellow tuber-
DEC+J
Model
DEC
cle at the angle of the jaw. It differs from A. latiffi

by lacking nuptial pads; having less webbing on

Figure 4. Dispersal–Extinction–Cladogenisis +J (DEC+J) chronogram depicting the most likely ancestral ranges for all
lineages in the genus Ansonia. Numbers at the nodes represent an estimate of the mean divergence time rounded to one
significant digit. The presence of Ansonia leptopus in Sumatra is in accordance with Frost et al. (2016).
toes I–III and V; having as opposed to lacking an on toes I and III; and having a coarsely granular
outer metatarsal tubercle; having as opposed to vs. a finely granular abdomen. These characters
lacking a dorsolateral row of enlarged tubercles on and others are scored across all other nominal spe-
the back; and lacking as opposed to having gular cies of Ansonia with the diagnostic differences of
spots and an interscapular spot. Ansonia each species highlighted (Table 4).
khaochangensis sp. nov. can be separated from Distribution. Ansonia khaochangensis sp. nov.
all Thai-Malay Peninsula species except Ansonia is known only from the type locality at Takua Pa
siamensis and A. latiffi by lacking as opposed to District, Phang Nga Province, southern Thailand
having submandibular tubercles in males. From A. (Fig. 1).
siamensis, it can be differentiated by the males Natural history. Ansonia khaochangensis sp.
being larger (maximum SVL = 32 mm vs. 28 mm); nov. is only known from the Khao Chang tower
having a visible tympanum; more extensive webbing karst formation that rises precipitously to an
sitting on the leaves of small plants growing out of

the karst. The distinctive, virtually monochromatic,
brown coloration of this species appears to be a
result of substrate matching in that frogs are reason-
ably well-camouflaged on the limestone substrate.
We interpret this as an indication that this micro-
habitat is not something that is occasionally utilized
and that this genus, which has a lotic life style else-
where, has adapted to this microhabitat over a long
period of time in that it split off from its sister lin-
eage approximately 5.5 Mya (Fig. 3).
The reproductive biology of this species is com-

pletely unknown. Most other Ansonia require fast-
moving lotic systems and all require water for tad-
pole development. We assume that there are other
sources of underground water that these toads must
be using for reproduction because there are no
above-ground systems nearby that we could find.
Fieldwork is currently being planned to investigate
this species’ presumably unique reproductive biology.
Etymology. The specific epithet khaochangensis is
in reference to the type locality of this species in the
Khao Chang tower karst formation.
DISCUSSION
Figure 5. Type series of Ansonia khaochangensis sp. H ISTORICAL BIOGEOGRAPHY OF THE GENUS ANSONIA
nov. from Khao Chang karst tower, Phang Nga, Takua Understanding the role that evolution has played in
Pa District, Phang Nga Province, Thailand. Upper: holo- the current composition of the megadiverse biota of
type ZMKU AM 01146. Lower: above from left to right, Sundaland is not possible without considering the
holotype ZMKU AM 01146, paratype ZMKU AM 01150, abiotic stage upon which tectonics and climate have
and paratype 12457; below from left to right, paratype performed over the last 50 Myr. From the middle
ZMKU AM 01148, paratype ZMKU AM 01149, and para- Eocene (approximately 42 Mya) onward, much of
type ZMKU AM 01145. what is today considered to be southern Sundaland
had subsided, leaving most of southern Borneo and
elevation of 125 m and frames the northwest border Java submerged beneath the South China Sea up
of the city of Phang Nga (Fig. 6). Along the southeast until the late Oligocene (approximately 25 Mya: de
face of the karst formation is the massive Phung Bruyn et al., 2014; Hall, 2013). However, as a result
Chang Cave that has a subterranean water system of the formation of upland areas in central Borneo
that runs beneath the karst tower. This southeast during the early Miocene (approximately 23 Mya),
face is also riddled with several smaller caves. this area remained land-positive and maintained a
Approximately 0.5 km southwest of the mouth of broad connection through the Thai-Malay Peninsula
Phung Chang Cave is a very small opening at the and the central Sunda Shelf with mainland Indo-
base of the karst tower that winds its way inward china, enabling everwet rainforest to expand north-
for approximately 20 m wherein we found A. ward (Morley, 2000, 2012). de Bruyn et al. (2014)
khaochangensis sp. nov. (Fig. 6). During the day, studied the phylogenetic structure of 61 Sundaic lin-
specimens were found only within the recesses of the eages of plants, invertebrates, and vertebrates and
cave on the vertical cave walls as high as 3 m above convincingly demonstrated that, as a result of Bor-
the cave floor. To escape, toads were able to move neo’s long history of being an upland refuge with a
sideways up the cave walls in an attempt to wedge generally stable climate, it served as a significant
themselves into narrow cracks near the celling. source and promotor of in situ speciation and the
Toads were also observed in the cave at night but predominant source of colonization for taxa emigrat-
would venture out to climb on the outside of the ing to other Sundaic landmasses, as well as the
karst tower. Some specimens were found as high as Philippines. Indeed, they demonstrated that, during
10 m above the forest floor both on the karst and the Miocene and Plio-Pleistocene (approximately 1.5–
Table 3. Selected characters states on the type series of Ansonia khaochangensis sp. nov. from Phang Nga, Takua
Pa District, Phang Nga Province, Thailand
ZMKU AM ZMKU AM ZMKU AM ZMKU AM ZMKU AM ZMKU AM

01146 01145 01147 01148 01149 01150
Holotype Paratype Paratype Paratype Paratype Paratype
Sex F F M F M F
Snout projecting beyond lower jaw (1) or not (0) Yes Yes Yes Yes Yes Yes
Tympanum visible (1) or not (0) Yes Yes Yes Yes Yes Yes
White or yellow wart at angle of jaw (1) or not (0) No No No No No No
Large yellow wart at angle of jaw (1) or not (0) No No No No No No
Interorbital tubercle ridges present (1) or not (0) No No No No No No

Opening of vocal sac on right (1) or left (0) / / Right / Right /
Finger tips rounded or forming small discs (1), 1 1 1 1 1 1
expanded or espatulate (0), or niether (0)
First finger reaching disc of second (1) or not (0) 0 0 0 0 0 0
Number of fingers with nuptual pads 0 0 0 0 0 0
Number of free phalages on fifth toe 2 2 2 2 2 2
Number of free phalages on fourth toe 3 3 3.5 3 3.5 3.5
Number of free phalages on third toe 2 2 2 2 2 2
Number of free phalages on second toe 1 1 1.5 1 1.5 1.5
Number of free phalages on first toe 0.5 0.5 0.5 0.5 0.5 0.5
Tarsal ridge present (1) or not (0) 1 1 1 1 1 1
Inner metatarsal tubercle (1) or not (0) 1 1 1 1 1 1
Outer metatarsal tubercle (1) or not (0) 1 1 1 1 1 1
Submandibular tubercles in males (1) or not (0) 0 0 0 0 0 0
Dorsal tubercles present (1) or not (0) 1 1 1 1 1 1
Dorsolateral row of enlarged tubercles present (1) 0 0 0 0 0 0
or not (0)
Oblique flap of skin on each side of vent (1) or 0 0 0 0 0 0
not (0)
Abdomen coarsley granular (1), finely granular (2), 1 1 1 1 1 1
or tuberculate (0)
Colour of iris Black Black Black Black Black Black
Gular spotting present (1) or not (0) 0 0 0 0 0 0
Wide, light patch below eye (1) or not (0) 0 0 0 0 0 0
Wide postorbital patch (1) or not (0) 0 0 0 0 0 0
Light spot between scapulae (1) or not (0) 0 0 0 0 0 0
Light crossbars on hind limbs (1) or not (0) 0 0 0 0 0 0
Vertebral stripe present (1) or not (0) 0 0 0 0 0 0
SVL 34.2 34.0 31.9 34.5 32.0 35.3
HL 10 9.8 9.2 9.9 9.4 10.6
HW 8.6 8.7 8.6 8.7 8.1 8.9
SL 4.2 4.1 4.0 4.2 4.2 4.4
SW 4.6 4.4 4.2 4.6 3.9 4.6
IND 1.9 2.1 1.9 2.0 1.8 2.1
ED 3.6 3.5 3.2 3.4 1.8 3.9
IOD 4.0 3.6 3.7 3.9 3.3 4.1
TD 2.1 2.1 2.2 2.0 1.8 2.1
CL 15.4 14.9 14.1 15.9 14.5 15.2
PL 11.8 11.8 10.5 12.3 10.9 11.1
F, female; M, male; SVL, snout–vent length; HL, head length from tip of snout to vent; head width, from posterior mar-
gin of mandible to tip of snout; SL, snout length, measured at the level of the jaw articulation; SW, snout width, mea-
sured from the anterior margins of the eyes to the tip of the snout; IND, internarial distance measured from the medial,
distance between the anterior margins of the eyes; ED, eye diameter, distance between the medial margins of the upper
eyelids at their closest point; IOD, interorbital diameter, inner margins of the nostrils; TD, tympanum diameter, the
length between anterior and posterior margins of the eye; CL, crus length, the length of the vertical axis; the distance
from the proximal edge of the outer metacarpal tubercle to the tip of the third finger; PL, pes length, the distance from
the knee inflection to the tarsal inflection.
Table 4. Diagnostic characters differentiating the species of Ansonia khaochangensis sp. nov. from all other nominal
species of Ansonia. Diagnostic characters are in bold text
Species albomaculata anotis endauensis fuliginea
SVL (female) 30–35 52.1 28.5 38–44

SVL (male) 20–28 37.1 17.4 22–36
Snout projecting beyond lower jaw (1) or not (0) 1 1 1 0
Tympanum visible (1) or not (0) 1 0 1 1
White or yellow wart at the angle 0 0 1 1
of the jaw present (1) or not (0)
Large yellow wart at angle of jaw present (1) or not (0) 0 0 0 0

Interorbital tubercle ridges present (1) or not (0) 0 0 0 0
Opening of vocal sac on right (1) or left (0) 1 or 0 Absent 1 and 0 1
Finger tips rounded or forming small discs (1) 1 0 1 1
or expanded and espatulate (0) or neither (2)
Toe tips rounded or forming small discs (1) 1 1 1 1
First finger reaching the disc of the second (1) or not (0) 0 0 0 0
Number of fingers with nuptial pads 1 0 0 1,2
Number of free phalanges of V toe 1 1–1.5 2 2
Number of free phalanges of IV toe 1.5–2 3 3.5 3
Number of free phalanges of III toe 1 1.5–2 2 1.5–2
Number of free phalanges of II toe 1 1.5–2 1 0.5–1
Number of free phalanges of I toe 1 1 1 0.5
Tarsal ridge present (1) or not (0) 1 0 0 0
Inner metatarsal tubercle present (1) or not (0) 1 1 1 0
Outer metatarsal tubercle present (1) or not (0) 1 1 1 1
Submandibular tubercles in males present (1) or not (0) 0 0 1 1
Dorsal tubercles present (1) or not (0) 1 1 1 1
Dorsolateral row of enlarged 0 0 0 0
tubercles present (1) or not (0)
Rows of tubercles on back (1) or not (0) 0 0 0 0
Oblique flap of skin on each side of vent (1) or not (0) 0 0 0 0
Abdomen coarsely grannular (1), 1 1 1 1
finely grannular (2), or tuberculate (0)
Colour of iris Gold / Red Gold
Gular spotting present (1) or not (0) 0 0 0 0
Wide, light patch below eye (1) or not (0) 1 0 0 0
White postorbital patch present (1) or not (0) 1 0 0 0
Light spot between the scapulae present (1) or not (0) 0 0 0 0
Light crossbars on hind limbs present (1) or not (0) 0 0 1 0
Vetebral stripe present (1) or not (0) / 1 1 and 0 1
23 Mya), more species from Borneo colonized the facilitated repeated range expansions during epi-
Philippines and the Thai-Malay Peninsula than from sodes of lowered sea levels followed by range retrac-
any other Asian landmass. Additionally, Pliocene tions and potential, vicariant speciation events with
and Quaternary climate and glacial-driven fluctua- their subsequent rise (Cannon et al., 2009; Gower
tions in sea levels ranged between 40 and 60 m et al., 2012; de Bruyn et al., 2013, 2014).
below present-day levels (Lohman et al., 2011). At The phylogenetic and biogeographical history of
their lowest levels, approximately 50% more terrene Ansonia aligns itself well with the overarching
of the Sunda Shelf was exposed than is exposed results outlined in de Bruyn et al. (2014) in that,
today (the smallest land-positive geographical area of subsequent to the early Miocene (approximately
the period) leaving the current Sundaic biota in a 23 Mya), Borneo served as an insular refuge, a
highly refugial state (Cannon, Morley & Bush, 2009; region of in situ speciation, and a source of coloniza-
Woodruff & Turner, 2009; Woodruff, 2010). These tion. Given that the current range of Ansonia
glacioeustatic events are considered to have extends throughout southwestern Indochina, most of
Table 4. Continued
Species glandulosa guibei hanitschi inthanon
SVL (Female) / 34 37 23.3–25.2

SVL (Male) 39.7 32 32 22.9–25.2
White or yellow wart at the angle / 1 1 /

Opening of vocal sac on right (1) or left (0) 1 Absent 1 1
Number of fingers with nuptial pads 1 1,2,3 1 1
Number of free phalanges of V toe 1.5 1 1.5–2 1
Number of free phalanges of IV toe 3 2.5 3 2.75
Number of free phalanges of III toe 1 1 1.5–2 0.5–2.66
Number of free phalanges of II toe 1 1 1 0.5–2
Number of free phalanges of I toe 1 1 1 0.5–1
Colour of iris / Gold Gold /
Gular spotting present (1) or not (0) / 0 0 1
Vetebral stripe present (1) or not (0) / 1 0 /
Sundaland, and the Philippines, suggests that its landmasses (Fig. 2) indicates that the phylogenetic
diversity and distribution was probably much greater structure within Ansonia and the current distribu-
on the exposed Sunda Shelf during any of its 50+ tion of its species are not the result of single vicari-
episodes of maximum exposure over the last 5 Myr ant episode and the reconstruction of the
when approximately 50% more terrene was land- biogeographical history of this genus’ major lineages
positive (Woodruff, 2010). Thus, extant, extra-Bor- could potentially be a reflection of multiple invasions
nean species of Ansonia merely represent refugial over broad expanses of time facilitated by the
lineages restricted to the continental fringes of the dynamic, episodic, physiogeographical history of Sun-
Sunda Shelf and, therefore, any fine-grained analysis daland (de Bruyn et al., 2014).
attempting to piece together their historical biogeog- The Dispersal–Extinction–Cladogenesis +J model
raphy in the absence of what are probably many, clearly indicates that Borneo is the ultimate source
extinct, more centrally located, basal lineages will of origin for all Thai, Peninsular Malaysian, Suma-
always be somewhat speculative. Nonetheless, the tran and Philippine populations of Ansonia (Fig. 4)
non-monophyly of suites of species on Sundaic in that all the basal lineages occur in this region.
Table 4. Continued
Species jeetsukumarani kraensis latidisca latiffi
SVL (Female) 25–25.3 24.0–27.9 55 50.7

SVL (Male) 19.4–19.9 19.9–22.3 35 34.5
White or yellow wart at the angle 1 1 / 0

Opening of vocal sac on right (1) or left (0) 1 0 0 1,0
Number of fingers with nuptial pads 1 1 0 0.5–1
Number of free phalanges of V toe 1.5–2 0.5 2 3
Number of free phalanges of IV toe 3 0.5–2.0 3.5 1
Number of free phalanges of III toe 1.5–2 0.5–2.33 2 1
Number of free phalanges of II toe 1 0.5–2.0 1 0–1
Number of free phalanges of I toe 0.5 0.5–1.0 1 1
Tarsal ridge present (1) or not (0) 1 or 0 0 0 1
Inner metatarsal tubercle present (1) or not (0) 1 or 0 1 1 1
Abdomen coarsely grannular (1), 1(M), 2(F) 1 1 2
Colour of iris Reddish/orange Gold / Reddish-gold
Vetebral stripe present (1) or not (0) 1 1 / 0
This pattern of Borneo being the source of origin for one major clade and a polyphyletic group in another
Sundaic lineages predominates other biotic groups as (Fig. 2).
well (de Bruyn et al., 2014). Lines of evidence point- At 13.0 Mya, an ancestral Bornean lineage
ing to Borneo as the centre of origin and diversifica- diverged into two major clades. By no later than
tion of Ansonia are that: (1) no species are known 11.7 Mya, one of these clades (clade 1) split into two
from Java, the southernmost island in the Sunda monophyletic groups, one of which remained in Bor-
Shelf that was submerged during much of the Mio- neo and began diversifying from approximately 9.2
cene when the uplands of Borneo were forming; (2) to at least 3.3 Mya and is represented currently by
Ansonia’s centre of diversity is on Borneo, 11 nomi- Ansonia platysoma, A. vidua, A. spinulifer, A. han-
nal species of at least 28 species as well as at least itschi, A. minuta, and at least two undescribed spe-
three undescribed species occur there; (3) the molec- cies (Fig. 4). The other monophyletic group of clade 1
ular analyses indicate that Bornean taxa form the gave rise to a monophyletic upland lineage that
ancestral lineages in the two major clades (Fig. 4); eventually invaded the southern portion of the Thai-
and (4) Bornean taxa form a paraphyletic group in Malay Peninsula south of the Isthmus of Kra no
Table 4. Continued
Species latirostra leptopus longidigita lumut
SVL (Female) 22.5–30.5 45–65 70 27.7–31.6

SVL (Male) 21.5–23.6 30–40 50 21–23.6
Interorbital tubercle ridges present (1) or not (0) 1 1,0 1,0 0

Opening of vocal sac on right (1) or left (0) 0 or 1 1 1 0
First finger reaching the disc 0 1 1 0
of the second (1) or not (0)
Number of fingers with nuptial pads 1 1 1–1,2 1
Number of free phalanges of V toe 1–2 2–2.5 1–2 <2
Number of free phalanges of IV toe 3–3.5 3–4 3 3.5–2
Number of free phalanges of III toe 1–2 2–2.5 0.5–2 1–3.5
Number of free phalanges of II toe 0.5–1 1 1 0.5–3
Number of free phalanges of I toe 0.5 1 1 1.5–2
Inner metatarsal tubercle present (1) or not (0) Very weak 0 1 1
Submandibular tubercles in males present (1) or not (0) 1 1 1 /
Colour of iris Gold-brown Gold Gold Dark-brown
Light spot between the scapulae present (1) or not (0) 0,1 0 0 0
Vetebral stripe present (1) or not (0) 1 0 0 0
later than 11.1 Mya (Figs 4, 7). Colonization was the southern Thai-Malay Peninsula giving rise to an
most likely by way of stream-capture-dispersal dur- upland and a generally lowland clade. The upland
ing this period of maximum everwet forest distribu- clade is currently composed of species endemic to
tion (de Bruyn et al., 2014) being that a wide land- montane regions of Peninsular Malaysia and con-
positive connection existed between Borneo and tains A. smeagol, Ansonia penangensis, A. lumut,
southern Indochina (Hall, 2013) and the climate was A. malayana, and A. jeetsukumaran. The lowland
relatively stable (Morley, 2000, 2012). This group is group dispersed northward out of the southern por-
currently represented by A. latidisca from the tion of the Thai-Malay Peninsula into southern Thai-
uplands of northwestern Borneo and a monophyletic land, crossing the Isthmus of Kra into the northern
group containing 15 species from the Thai-Malay portion of the Thai-Malay Peninsula (southern Indo-
Peninsula. Between 9.7 and 11.1 Mya, the Thai- china) no later than 6.7 Mya (Figs 4, 7). Dispersal as
Malay lineage of this group separated from the Bor- opposed to vicariance is invoked for this event given
nean lineage and, by 9.7 Mya, began diversifying in that the species south of the Isthums of Kra are
Table 4. Continued
Species malayana mcgregori minuta muelleri
SVL (Female) 24–29 43–50 29 30–38

SVL (Male) 20–23 32–39 20–24 25–31
Large yellow wart at angle 1 0 0 0
of jaw present (1) or not (0)

Opening of vocal sac on right (1) or left (0) 1 0 or 1 0 0 or 1
First finger reaching the disc 0 0 0 0
of the second (1) or not (0)
Number of fingers with nuptial pads 1 1 0 1
Number of free phalanges of V toe 1 1 1 1
Number of free phalanges of IV toe 4 1–2.5 2 1–3
Number of free phalanges of III toe 1 1 1 1
Number of free phalanges of II toe 0–1 1 1 1
Number of free phalanges of I toe 0–1 1 1 1
Submandibular tubercles in 1 0 1 1
males present (1) or not (0)
Dorsal tubercles present (1) or not (0) 1 1,2 1,2 1,2
Colour of iris Gold / / /
Vetebral stripe present (1) or not (0) 0,1 0 1 /
paraphyletic with respect to those north of the Isth- of these lineages, currently composed of species found
mus of Kra. A vicariant event of that age would have primarily in northeastern Borneo (Ansonia fuliginea
most likely resulted in two reciprocally monophyletic and Ansonia guibei), invaded the Philippines early on
groups. By at least 5.5 Mya, the monophyletic (approximately 8.1 Mya) via the Sulu-Mindanao
Indochinese lineage began diversifying and eventu- Archipelago (Sanguila et al., 2011). Sanguila et al.
ally gave rise to A. khaochongensis sp. nov., Anso- (2011) proposed the mode of dispersal through the
nia sp. 4 and 5., A. inthanon, A. thinthinae, and archipelago was by island hopping, which appears to
A. kraensis (Figs 3, 4). be inconsistent with this group’s lotic habitat require-
The other major Bornean clade (clade 2) gave rise ment but, given there was no continuous land-posi-
to two lineages no later than 10.0 Mya (Fig. 4). One tive connection between Borneo and Mindanao
Table 4. Continued
Species penangensis platysoma siamensis smeagol
SVL (Female) 37.2 20–26 35 24.6–27.4

SVL (Male) / 20–25 28 20

Opening of vocal sac on right (1) or left (0) / 0 0 or 1 0
or expanded and estapulate (0) or neither (2)
First finger reaching the disc of the 0 1 0 0
second (1) or not (0)
Number of fingers with nuptial pads / 1 1 /
Number of free phalanges of V toe 1.5 1–1.5 1 1.5
Number of free phalanges of IV toe 3 3 2 3.5–3.7
Number of free phalanges of III toe 1.5 1 1 1.5–2
Number of free phalanges of II toe 0 1 1 0.5–2
Number of free phalanges of I toe 0 1 1 1
Tarsal ridge present (1) or not (0) 0,1 0 0 0
Submandibular tubercles / 0 0 /
in males present (1) or not (0)
Dorsolateral row of enlarged tubercles 0 0 0 0
present (1) or not (0)
Colour of iris / Gold / Dark w/gold
specks
Vetebral stripe present (1) or not (0) 1 0 0 1
during this time period (Blackburn et al., 2010; Siler The other lineage of clade 2 began diversifying in
et al., 2012; Hall, 2013), this hypothesis is followed Borneo no later than 9.0 Mya and eventually gave
here. Once established, this lineage began diversify- rise to a upland group composed of Ansonia albomac-
ing by at least 3.7 Mya following the uplift of the ulata and Ansonia leptopus and a related group of
Mindanao highlands, giving rise to the Ansonia muel- similar species designated here to form a species com-
leri complex and its sister species Ansonia mcgregori plex. By 5.0 Mya, the latter began diversifying into
(Sanguila et al., 2011). Although the age of this clado- what is now a complex of species in dire need of taxo-
geneic event is older than that proposed by Sanguila nomic study. Recently, one lineage of the leptopus
et al. (2011; approximately 2.2 Mya), the error bars complex entered Peninsular Malaysia by no later
of both time-calibrated BEAST trees overlap widely. than 1.1 Mya, probably during one of the many
Table 4. Continued
Species spinulifer tiomanica torrentis khaochangensis
SVL (Female) 50 38.4 / 35.5

SVL (Male) 41 31.2 30–33 32
White or yellow wart at the angle 1 1 / 0

Opening of vocal sac on right (1) or left (0) 1 1 0 or 1 1
Number of fingers with nuptial pads 1 0 1 0
Number of free phalanges of V toe No web 2 2 2
Number of free phalanges of IV toe No web 3.5 3.5–3.7 3–3.5
Number of free phalanges of III toe No web 2 2 2
Number of free phalanges of II toe No web 1 / 1–1.5
Number of free phalanges of I toe No web 1 / 0.5
Submandibular tubercles in 1 1 1 0
males present (1) or not (0)
Dorsolateral row of enlarged tubercles 1 0 0 1
present (1) or not (0)
Colour of iris Gold / / Black
Light spot between the 1 0 0 0
scapulae present (1) or not (0)
Light crossbars on hind 0 1 1 0
limbs present (1) or not (0)
Vetebral stripe present (1) or not (0) 0 1 / 0
/, data unavailable. SVL, snout–vent length.

Bold character states are those that discretely separate A. khaochangensis sp. nov. from the species in the respective
column.
episodes when lower sea levels produced a land-posi- vicinity of 0.84 Mya (Fig. 3), this continuous popula-
tive connection between Borneo and Peninsular tion became restricted to separate, upland regions.
Malaysia and lowered temperatures allowed upland This separation is represented today by A. latiffi in
forests and their associated riparian habitats to Peninsular Malaysia. Similarly, by approximately
migrate downslope and expand across the interven- 2.0 Mya, another lineage of the leptopus complex
ing lowlands (Cannon et al., 2009). When higher tem- diversified and subsequently invaded Sumatra
peratures and higher sea levels returned in the (Figs 4 and 7).
Figure 6. Upper: the karst tower Khao Chang at the type locality. Lower left: entrance to the small cave where speci- Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022
mens the type series were collected. Lower right: interior of the cave where specimens were collected.
C ONSERVATION assume that species evolution in Borneo is a phe-

The Bornean highlands are a series of mountains nomenon restricted to only the past. Indeed, the
harboring a remarkably high number of species data for Ansonia indicate that some of these specia-
and have long served as a source of biodiversity for tion events have happened in as little as 0.34 Mya
other Southeast Asian regions (de Bruyn et al., (Fig. 3). Therefore, conservation in Borneo should
2014). Speciation in Borneo has been an ongoing be a high priority not only to protect current lin-
process subsequent to the Miocene (de Bruyn et al., eages, but also to maintain this island as a refuge
2014) and evolution within the genus Ansonia fits area for the entire Sunda Shelf and future
this model well. However, there is no reason to lineages.

Figure 7. Map showing the out-of-Borneo scenario for the evolution of major lineages of Ansonia and their spread
throughout Sundaland and the Philippines. Arrows indicate the origin and direction of colonization only, and not the
mode of colonization. Distribution ellipses denote the presence or absence of a lineage on a landmass, and not population
size or distribution.
Grismer et al. (2016, 2014) discussed the rich, departure from the typical reproductive biology of
underappreciated, and understudied herpetological this genus assuming breeding is not taking place in
diversity of karst ecosystems throughout Southeast subterranean bodies of water. More importantly,
Asia west of Wallace’s Line. These discussions, how- however, this highlights the fact that not only do
ever, pertained to reptiles, most notably gekkonids, karst ecosystems promote the evolution of new spe-
whose scansorial proclivities somewhat preadapt cies, but also they can contribute to the evolution of
them to these microhabitats. The discovery of a new notably different life styles.
species of karst-dwelling anuran in Southeast Asia is
not unprecedented (Siler et al., 2010), although it is
the first such record from the Thai-Malay Peninsula.
ACKNOWLEDGEMENTS
Of more probable significance, however, is the repro-
ductive biology of this new species. The karst-dwell- Funding for LLG came from grants from the College
ing species described by Siler et al. (2010) of the of Arts and Sciences at La Sierra University and from
genus Platymantis from central Luzon Island in the a National Geographic Society Explorers Grant (9277-
Philippines, probably have direct-developing eggs, as 15). Partial funding for PLWJ and CA was from a NSF
has been reported for other species of Platymantis dimensions grant EF-1241885 issued to Jack W. Sites
that have been observed to deposit their eggs in Jr (JWSJ). Additional funding for PLWJ was from a
limestone crevices. Ansonia, however, do not lay Doctoral Dissertation Improvement Grant (DDIG
direct-developing eggs and the majority of the species 1501198) issued to PLWJ and JWSJ. We wish to
are restricted to lotic environments. From what we thank two anonymous reviewers and Masafumi Mat-
can surmise so far, this represents a significant sui for many helpful comments.
REFERENCES new Stream Toad of the genus Ansonia Stoliczka, 1870

(Anura: Bufonidae) from a montane region in Peninsular
Berry PY. 1975. The amphibian fauna of peninsular Malay- Malaysia. Zootaxa 4103: 137–153.
sia. Kuala Lumpur: Tropical Press, 130. Drummond AJ, Ashton B, Buxton S, Cheung M, Cooper
Blackburn DC, Bickford DP, Diesmos AC, Iskandar DT, A, Duran C. 2011. Geneious V5.4. Available at: http://
Brown RM. 2010. An ancient origin for the enigmatic www.geneious.com/
Flat-Headed Frogs (Bombinatoridae: Barbourula) from the Drummond AJ, Suchard MA, Xie D, Rambaut A. 2012.
islands of Southeast Asia. PLoS ONE 5: 1–10. Bayesian phylogenetics with BEAUti and BEAST 1.7.
van Boxclear I, Biju SD, Loader SP, Bossuyt F. 2009. Molecular Biology and Evolution 29: 1969–1973.
Toad radiation reveals into-India dispersal as a source of Edgar R. 2004. MUSCLE: multiple sequence alignment with
endemism in the Western Ghats-Sri Lanka biodiversity hot- high accuracy and high throughput. Nucleic Acids Research
spot. BMC Evolutionary Biology 9: 1–10. 32: 1792–1797.
Brandley MC, Wang Y, Guo X, Nieto Montes de Oca A,

Frost DR, Faivovich GT, Bain R, Haas A, Haddad
Feria Ortiz M, Hikida T, Ota H. 2011. Accommodating CFB, de Sa RO, Channing A, Wilkinson M, Donnel-
locus-specific heterogeneity in molecular dating methods: lan SC, Raxworthy C, Campbell JA, Blotto BL,
an example using inter-continental dispersal of Plestiodon Moler P, Drewes RC, Nussbaum RA, Lynch JD,
(Eumeces) lizards. Systematic Biology 60: 3–15. Green DM, Wheeler WC. 2016. The amphibian tree of
de Bruyn M, R€ uber L, Nylinder S, Stelbrink B, Lovejoy life. Bulletin of the American Museum of Natural History
NR, Lavou� e S, Tan HH, Nugroho E, Wowor D, Ng 297: 1–1291.
PKL, Siti Azizah MN, von Rintelen T, Hall R, Car- Gower D, Johnson KG, Rosen BR, Richardson J, R€ uber
valho GR. 2013. Paleo-drainage basin connectivity predicts L, Williams ST. 2012. Biotic evolution and environmental
evolutionary relationships across three Southeast Asian bio- change in Southeast Asia. Cambridge: Cambridge Univer-
diversity hotspots. Systematic Biology 62: 398–410. sity Press.
de Bruyn M, Stelbrink B, Morley RJ, Hall R, Carvalho Grandison AGC. 1972. The gunong benom expedition 1967.
GR, Cannon CH, van den Bergh G, Meijaard E, Met- 5. Reptiles and amphibians of Gunong Benom with a
calfe I, Boitani L, Maiorano L, Shoup R, von Rintelen description of a new species of Macrocalamus. Bulletin of
T. 2014. Borneo and Indochina are major evolutionary hot- the British Museum of Natural History (Zoology) 23: 45–
spots for southeast Asian biodiversity. Systematic Biology 101.
63: 1–23. Graybeal A. 1997. Phylogenetic relationships of bufonid
Cannon CH, Morley RJ, Bush ABG. 2009. The current frogs and tests of alternate macroevolutionary hypotheses
refugial rainforests of Sundaland are unrepresentative of characterizing their radiation. Zoological Journal of the
their biogeographic past and highly vulnerable to distur- Linnean Society 119: 297–338.
bance. Proceedings of the National Academy of Sciences of Grismer LL. 2011. Field guide to the amphibians and rep-
the United States of America 106: 11188–11193. tiles of the seribuat archipelago (Peninsular Malaysia).
Carranza S, Arnold EN, Mateo JA, Lopez-Jurado LF. Franfurt am Main: Edition Chimaira. 239 pp.
2000. Long-distance colonization and radiation in gekkonid Grismer LL, Wood PL Jr, Anuar S, Quah ESH, Muin
lizards, Tarentola (Reptilia: Gekkonidae), revealed by MA, Chan KO, Sumarli AX, Loredo AL. 2015. Repeated
mitochondrial DNA sequences. Proceedings of the Royal evolution of sympatric, paleoendemic species in closely
Society of London Series B, Biological Sciences 267: 637– related, co-distributed lineages of Hemiphyllodactylus Blee-
649. ker, 1860 (Squamata: Gekkonidae) across a sky-island
Chan KO, Wood PL Jr, Anuar S, Muin MA, Quah ESH, archipelago in Peninsular Malaysia. Zoological Journal of
Sumarli AX, Grismer LL. 2014. A new species of upland the Linnean Society 174: 859–876.
Stream Toad of the genus Ansonia Stoliczka (Aunra: Bufo- Grismer LL, Wood PL Jr, Anuar S, Quah ESH, Muin
nidae) from northeastern Peninsular Malaysia. Zootaxa MA, Mohamed M, Chan KO, Sumarli AX, Loredo AI,
3764: 427–440. Heinz HM. 2014. The phylogenetic relationships of three
Crawford AJ. 2003a. Huge populations and old species of new species of the Cyrtodactylus pulchellus complex (Squa-
Costa Rican and Panamanian dirt frogs inferred from mito- mata: Gekkonidae) from poorly explored regions in north-
chondrial and nuclear gene sequences. Molecular Ecology eastern Peninsular Malaysia. Zootaxa 3786: 359–381.
12: 2525–2540. Grismer LL, Wood PL Jr, Anuar S, Davis HR, Cobos
Crawford AJ. 2003b. Relative rates of nucleotide substi- AJ, Murdoch ML. 2016. A new species of karst forest
tution in frogs. Journal of Molecular Evolution 57: 636– Bent-toed Gecko (genus Cyrtodactylus Gray) not yet threat-
641. ened by foreign cement companies and a summary of
Darst CR, Cannetella DC. 2004. Novel relationships among Peninsular Malaysia’s endemic karst forest herpetofauna
hyloid frogs inferred from 12S and 16S mitochondrial DNA and the need for its conservation. Zootaxa 4061: 1–17.
sequences. Molecular Phylogenetics and Evolution 31: 462– Grummer JA, Bryson RW Jr, Reeder TW. 2014. Species
475. delimitation using Bayes factors: simulations and applica-
Davis HR, Grismer LL, Wood PL Jr, Muin MA, Anuar tion to the Sceloporus scalaris species group (Squamata:
S, Klabacka R. 2016. The phylogenetic relationships of a Phrynosomatidae). Systematic Biology 63: 119–133.
Hall R. 2013. The palaeogeography of Sundaland and Wal- phylogenetic trees. In: Proceedings of the Gateway Comput-
lacea since the Late Jurassic. Journal of Limnology 72: 1– ing Environments workshop (GCE), New Orleans, LA, 1–8.
17. Minh Q, Nguyen MAT, von Haeseler A. 2013. Ultrafast
Heinicke MP, Greenbaum E, Jackman TR, Bauer AM. approximation for phylogenetic bootstrap. Molecular Biol-
2011. Phylogeny of a trans- Wallacean radiation (Squa- ogy and Evolution 30: 1188–1195.
mata, Gekkonidae, Gehyra) supports a single early coloniza- Morley RJ. 2000. Origin and evolution of tropical rain for-
tion of Australia. Zoologica Scripta 40: 584–602. ests. Chichester: John Wiley & Sons.
Hertwig ST, Min PY, Haas A, Das I. 2014. Dressed in Morley RJ. 2012. A review of the Cenozoic palaeoclimate
black. A New Ansonia Stoliczka, 1870 (Lissamphibia: history of Southeast Asia. In: Gower D, Johnson KG, Rosen
Anura: Bufonidae) from Gunung Murud, Sarawak, East BR, Richardson J, R€ uber L, Williams ST, eds. Biotic evolu-
Malaysia (Borneo). Zootaxa 3814: 419–431. tion and environmental change in Southeast Asia. Cam-
Huelsenbeck J, Ronquist F. 2001. MRBAYES: Bayesian bridge: Cambridge University Press, 79–114.

inference of phylogeny. Bioinformatics 17: 754–755. Nguyen L-T, Schmidt HA, von Haeseler A, Minh BQ.
Inger RF, Stuebing RB. 2005. A field guide to the frogs of 2015. IQ-TREE: a fast and effective stochastic algorithm for
Borneo. Kuching: Natural History Publications (Borneo), estimating maximum likelihood phylogenies. Molecular
201. Biology and Evolution 32: 268–274.
Landis M, Matzke NJ, Moore BR, Huelsenbeck JP. Rambaut A, Drummond AJ. 2013. TreeAnnotator, Version
2013. Bayesian analysis of biogeography when the number 1.8.0. MCMC output analysis. Available at: http://beast.-
of areas is large. Systematic Biology 62: 789–804. bio.ed.ac.uk/TreeAnnotator
Lohman DJ, de Bruyn M, Page T, von Rintelen K, Hall Rambaut A, Suchard MA, Xie D, Drummond AJ. 2014.
R, Ng PKL, Shih HT, Carvalho GR, von Rintelen T. Tracer, Version 1.6. Available at: http://beast.bio.ed.ac.uk/
2011. Biogeography of the Indo-Australian archipelago. Tracer
Annual Review of Evolution, Ecology and Systematics 42: Ree RH, Smith SA. 2008. Maximum likelihood inference of
205–226. geographic range evolution by dispersal, local extinction,
Macey JR, Schulte JA II, Larson A, Fang Z, Wang Y, and cladogenesis. Systematic Biology 57: 4–14.
Tuniyev BS, Papenfuss TJ. 1998. Phylogenetic relation- Ronquist F. 1997. Dispersal–vicariance analysis: a new
ships of toads of the Bufo bufo species group from the east- approach to the quantification of historical biogeography.
ern escarpment of the Tibetan Plateau: a case of vicariance Systematic Biology 46: 195–203.
and dispersal. Molecuar Biology and Evolution 9: 80–87. Ronquist F, Teslenko M, van der Mark P, Ayres DL,
Macey JR, Strasburg JL, Brisson JA, Vredenburg VT, Darling A, H€ ohna B, Larget L, Liu L, Suchard MA,
Jennings M, Larson A. 2001. Molecular phylogenetics of Huelsenbeck JP. 2012. Mr. Bayes 3.2: efficient Bayesian
western North American frogs of the Rana boylii species phylogenetic inference and model choice across a large
group. Molecular Phylogenetics and Evolution 19: 131–143. model space. Systematic Biology 61: 539–542.
Matsui M, Yambun P, Sudin A. 2007. Taxonomic relation- Sanguila MB, Siler DM, Diesmos AC, Nu~ nez O, Brown
ships of Ansonia anotis Inger, Tan, and Yambun, 2001 and RM. 2011. Phylogeography, geographic structure, genetic
Pedostibes maculatus (Mocquard, 1890), with a description variation, and potential species boundaries in Philippine
of a new genus (Amphibia, Bufonidae). Zoological Science 2: slender toads. Molecular Phylogenetics and Evolution 61:
1159–1166. 333–350.
Matsui M, Tominaga A, Liu W-Z, Khonsue W, Grismer Savage JM, Heyer WR. 1997. Digital webbing formulae for
LL, Diesmos AC, Das I, Sudin A, Yambun P, Yong H- Anurans: a refinement. Herpetological Review 28: 131.
S, Sukumaran J, Brown RM. 2010. Phylogenetic rela- Siler CD, Diesmos AC, Linkem CW, Diesmos ML, Brown
tionships of Ansonia from Southeast Asia inferred from RM. 2010. A new species of limestone-forest frog, genus
mitochondrial DNA sequences: systematic and biogeo- Platymantis (Amphibia: Anura: Ceraobatrachidae) from cen-
graphic implications (Anura: Bufonidae). Molecular Phylo- tral Luzon Island, Philippines. Zootaxa 2482: 49–63.
genetics and Evolution 54: 561–570. Siler CD, Oaks JR, Welton LJ, Linkem CW, Swab JJ, Dies-
Matsui M, Nishikawa K, Siew TY, Eto K. 2012. Notes on mos A, Brown RM. 2012. Did geckos ride the Palawan raft to
a rare Bornean bufonid Ansonia latidisca Inger, 1966, with the Philippines? Journal of Biogeography 39: 1217–1234.
special reference to is phylogenetic position. Current Her- Tan AM, Wake DB. 1995. MtDNA phylogeography of the
petology 31: 87–96. California newt, Taricha torosa (Caudata, Salimandridae).
Matzke NJ. 2013. Probabilistic historical biogeography: new Molecular Phylogenetics and Evolution 4: 383–394.
models for founder-event speciation, imperfect detection, Wang IJ, Crawford AJ, Bermingham E. 2008. Phylogeog-
and fossils allow improved accuracy and model-testing. raphy of the pygmy rain frog (Pristamantis ridens) across
Frontiers in Biogeography 5: 242–248. the lowland wet forests of isthmian Central America. Molec-
Matzke NJ. 2014. Model slection in historical biogeography ular Phylogenetics and Evolution 47: 992–1004.
reveals that founder-effect speciation is a crucial process in Wilcox TP, Zwickl DJ, Heath TA, Hillis DM. 2002.
island clades. Systematic Biology 63: 1–21. Phylogenetic relationships of the Dwarf Boas and a compar-
Miller MA, Pfeiffer W, Schwartz T. 2010. Creating the ison of Bayesian and bootstrap measures of phylogenetic sup-
CIPRES Science Gateway for inference of large port. Molecular Phylogenetics and Evolution 25: 361–371.
Wilkinson JA, Sellas AB, Vindum JV. 2012. A new species Woodruff DS, Turner LM. 2009. The Indochinese-Sundaic
of Ansonia (Anura: Bufonidae) from northern Tanintharyi zoogeographic transition: a description and analysis of ter-
Division, Myanmar. Zootaxa 3163: 54–68. restrial mammal species distributions. Journal of Biogeog-
Woodruff DS. 2010. Biogeography and conservation in raphy 36: 803–821.
Southeast Asia: how 2.7 million years of repeated environ- Zamudio KR, Jones KB, Ward RH. 1997. Molecular sys-
mental fluctuations affect today’s patterns and the future of tematicsof short-horned lizards: biogeography and taxon-
the remaining refugial-phase biodiversity. Biodiversity Con- omy of a widespread species complex. Systematic Biology
servation 19: 919–941. 46: 284–305.

Out of Borneo - Biogeography Katak

Uploaded by

Copyright:

Available Formats

Out of Borneo - Biogeography Katak

Uploaded by

Document Information

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

Out of Borneo - Biogeography Katak

Uploaded by

Copyright:

Available Formats

Biological Journal of the Linnean Society, 2017, 120, 371–395. With 7 figures.

Out of Borneo, again and again: biogeography of the

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

KEYWORDS: BioGeoBEARS – biogeography – karst ecosystem – Sundaland – systematics – Thailand.

INTRODUCTION the focus of biogeographers studying the history of

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

snout and canthus sparsely covered with small,

tuberculate; back bearing low, small tubercles; large

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

manus weakly tuberculate; forelimbs and fingers

proximal subarticular tubercles; fingertips rounded,

slightly dilated but not forming discs; subarticular

tubercles distinct; outer palmar tubercle oval, raised;

webbing formula: I 1, II ½–2, III 1½–2, IV 2–2, V 1;

ular tubercles weak; inner metatarsal tubercle elon-

gate, raised; outer metatarsal tubercle raised, oval,

colour of flanks, forelimbs, hands, and feet dull-

flanks, and fewer yellowish tubercles on limbs and

the holotype in all aspects of coloration and morphol-

as opposed to being dull-orange. Measurements

parisons were obtained from the data matrix of

Davis et al. (2016). Ansonia khaochangensis sp.

posed of 14 other species of Ansonia endemic to the

the hind limbs and having a black iris as opposed

to other colours (Table 4). It can be differentiated

from all the Thai-Malay species except Ansonia lat-

cle at the angle of the jaw. It differs from A. latiffi

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

sitting on the leaves of small plants growing out of

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

ZMKU AM ZMKU AM ZMKU AM ZMKU AM ZMKU AM ZMKU AM

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Species albomaculata anotis endauensis fuliginea

SVL (female) 30–35 52.1 28.5 38–44

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Species glandulosa guibei hanitschi inthanon

SVL (Female) / 34 37 23.3–25.2

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Species jeetsukumarani kraensis latidisca latiffi

SVL (Female) 25–25.3 24.0–27.9 55 50.7

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Species latirostra leptopus longidigita lumut

SVL (Female) 22.5–30.5 45–65 70 27.7–31.6

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Species malayana mcgregori minuta muelleri

SVL (Female) 24–29 43–50 29 30–38

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Species penangensis platysoma siamensis smeagol

SVL (Female) 37.2 20–26 35 24.6–27.4

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022

Species spinulifer tiomanica torrentis khaochangensis

SVL (Female) 50 38.4 / 35.5

Downloaded from https://academic.oup.com/biolinnean/article/120/2/371/2954941 by guest on 05 February 2022