A new karst-dwelling bent-toed gecko (Squamata: Gekkonidae: Cyrtodactylus) from Xiangkhoang Province, northeastern Laos

2018·20.4 MB·

by  Nazarov, Roman A

ZOOLOGICAL RESEARCH
A new karst-dwelling bent-toed gecko (Squamata:
Gekkonidae: Cyrtodactylus) from Xiangkhoang
Province, northeastern Laos
RomanA.Nazarov1,*,OlivierS.G.Pauwels2,EvgeniyL.Konstantinov3,AnatoliyS.Chulisov3,NikolaiL.Orlov4,
NikolayA.Poyarkov,Jr.5,6,*
1ZoologicalMuseum,MoscowStateUniversity,Moscow125009,Russia
2RoyalBelgianInstituteofNaturalSciences,BrusselsB-1000,Belgium
3InstituteofNaturalSciences,KalugaStateUniversitynamedafterK.I.Tsiolkovskii,Kaluga248023,Russia
4ZoologicalInstitute,RussianAcademyofSciences,St.Petersburg199034,Russia
5DepartmentofVertebrateZoology,BiologicalFaculty,LomonosovMoscowStateUniversity,Moscow119234,Russia
6JointRussian-VietnameseTropicalResearchandTechnologicalCenter,63NguyenVanHuyenRoad,NghiaDo,CauGiay,Hanoi,Vietnam
ABSTRACT (Nguyenetal.,2017). AlthoughLaosislocatedbetweenthese
twospecies-richcountriesandharborsavariedgeologicalrelief
Wedescribeanewkarst-dwellingCyrtodactylus from
propitioustomicro-endemism,itsCyrtodactylusfaunaiscurrently
Ban Thathom, Xiangkhoang Province, northeastern
knowntoincludeonly21species(Luuetal.,2016a;Nazarovet
Laos. The new species can be distinguished from
al., 2014). Such low species number may be due to the fewer
other congeners by having four dark dorsal bands herpetologicalsurveysconductedinLaoscomparedtoVietnam
betweenlimbinsertions,adiscontinuousnuchalloop, and Thailand, particularly on the karst reliefs (Teynié & David,
10precloacalporesinmalesor10–12precloacalpits 2014).
Our team co-described five of the Cyrtodactylus species
(females) separated by a diastema from a series of
currently recorded from Laos (Nazarov et al., 2014; Ngo &
enlargedfemoralscalesbearing18or19pores(male)
Pauwels, 2010), and we are pursuing our efforts to better
or8–10pits(females)alongeachfemur,14–18dorsal
inventorythediversityofthegenusinthiscountry.
tuberclerowsatmidbody,noprecloacalgroove,30–36 DuringfieldworkintheforestedkarstmassifsintheLaotian
midbodyscalerowsacrossbellybetweenventrolateral provinceofXiangkhoang,we(A.S.C.andE.L.K.)encountered
skinfolds,transverselyenlargedsubcaudalplates,and three geckos that differed in color pattern and scalation from
known species, and which we show hereafter to belong to
amaximalknownsnout-ventlengthof75.5mm. Our
anundescribed species. Morphological characters, such as
descriptionbringsto22thenumberofCyrtodactylus
speciesrecordedfromLaos.
Keywords: Indochinese region; Karst; Limestone; Received: 03September2017;Accepted: 19March2018;Online: 20
Herpetology;Taxonomy;Newspecies April2018
Foundation items: The work was partially supported by the
INTRODUCTION
Russian Foundation of Basic Research (RFBR 15-04-08393; RFBR
ThediversityofthegeckogenusCyrtodactylushasincreasedat 15-29-02771; RFBR 17-54-54002_viet_a; RFBR 18-04-00040);
a phenomenal rate. Of the more than 250 species recognized Preservationandstorageofspecimenswascarriedoutwithfinancial
withinthegenus,about80%ofthediversityhasbeendescribed support from the Russian Science Foundation (RSF 14-50-00029).
in the twenty-first century (Uetz et al., 2018). Numbers of
TheresearchwascompletedwithintheframeworksofRussianState
Cyrtodactylus species in Thailand and Vietnam have shown
projectsAAAA-A16116021660077-3andAAAA-A17-117030310017-8
considerableexpansion,includingatleast33and39recognized
species in each country, respectively (Pauwels et al., 2016). *Corresponding authors, E-mail: [email protected]; n.poyarkov@
Moreover, several species remain to be named in Vietnam, in gmail.com
particular within the Cyrtodactylus irregularis species complex DOI:10.24272/j.issn.2095-8137.2018.010
202 SciencePress ZoologicalResearch39(3):202–219,2018
scalation and dorsal color and pattern, indicate that this Morphologicaldescriptions
newly discovered Cyrtodactylus population clearly belongs to
Measurements and meristic counts follow Pauwels et al.
the C. phongnhakebangensis species group sensu Luu et
(2016). Measurements were taken on the right side. Paired
al. (2016a), which includes a dozen species from Laos and
meristic characters are given in left/right order. Numbers
Vietnam.Inagreementwiththedefinitionofthisgroupprovided
of supralabial and infralabial scales were counted from the
by Luu et al. (2016a: 132), this new species exhibits a
largest scale immediately posterior to the dorsal inflection of
maximum adult SVL comprised between 73.0 and 100.6 mm,
theposteriorportionoftheupperjawtotherostralandmental
0 or 1 supranasal, a DorTub (14–18) between 10 and 24, no
scales, respectively; the number of longitudinal rows of body
webbing between fingers or toes, tubercles nearly absent on
tubercles was counted transversely across the center of the
forelimbs but present on hind limbs, precloacal and femoral
dorsumfromoneventrolateralskinfoldtotheother;thenumber
pores in males (in total 47) between 20 and 60, number
oflongitudinalrowsofventralscaleswascountedtransversely
of postcloacal tubercles (5 or 6 in male) between 3 and 8,
acrossthecenteroftheabdomenfromoneventrolateralskin
enlargedsubcaudals,anddorsumdisplayingwell-defineddark
fold to the other; subdigital lamellae beneath the toes were
bands.
countedfromthebaseof thefirstphalanxtotheclaw; dorsal
MATERIALSANDMETHODS darkbandsbetweenlimbinsertionsarethosestrictlycontained
on the dorsum between the posterior insertion of the anterior
Sampling limbs and the anterior insertion of the posterior limbs. In
Field surveys were conducted in Xiangkhoang Province, previousliterature,dorsalbandsoftenincludethosecontained
Laos, from May to July 2008 within the frameworks of the between the anterior insertion point of the anterior limbs and
Memorandum of Understanding (MOU) on the Academic and the posterior point of insertion of the posterior limbs, thus
ScientificCooperationbetweentheNationalUniversityofLaos interspecific comparisons on band numbers were performed
andKalugaStateUniversity,Russia(MOUNo.7220-11-2012). withcaution.
Specimens were collected by hand from 1900 h to 2300 Thefollowingmeasurementsweretakenwithadigitalcaliper
h. Specimens were photographed in life and subsequently to the nearest 0.1 mm: AG: axilla to groin length, taken from
euthanized in a closed vessel with a piece of cotton wool theposteriormarginoftheforelimbatitsinsertionpointonthe
containing ethyl acetate (Simmons, 2002). Tissue samples body to the anterior margin of the hind limb at its insertion
were preserved separately in 95% ethanol and specimens point on the body; EarL: ear length, the greatest horizontal
were fixed in 85% ethanol, after which they were transferred distance of the ear opening; ForeaL: forearm length, taken
to 70% ethanol for permanent storage. Specimens were on the dorsal surface from the posterior margin of the elbow
subsequently deposited in the herpetological collection of the while flexed 90◦ to the inflection of the flexed wrist; HeadH:
ZoologicalInstituteoftheRussianAcademyofScience(ZISP) head height, the maximum height of head from the occiput
andintheherpetologicalcollectionoftheZoologicalMuseum to the throat; HeadL: head length, from the posterior margin
of Lomonosov Moscow State University (ZMMU), Moscow, of the retroarticular process of the lower jaw to the tip of the
Russia. snout;HeadW:headwidth,measuredattheangleofthejaws;
The specimens accessed for molecular analyses are Internar: internarial distance, measured between the nares
listed in Table 1. Our sampling is to date the most across the rostrum; Interorb: interorbital distance, measured
comprehensivesamplingcoveringallknownlineagesoftheC. between the anterior edges of the orbits; ML: mental length,
phongnhakebangensis species complex. A map showing the the maximum length of mental shield; MW: mental width, the
distribution of the C. phongnhakebangensis species complex maximum width of mental shield; NosOrb: nostril to orbit
in Laos and adjacent territories and the location of the distance,fromtheposteriormarginoftheexternalnarestothe
sampling site for the present work is provided in Figure anteriormarginoftheorbit;OrbD:orbitdiameter,thegreatest
1. Museum abbreviations used are listed as follows: CAS: horizontaldiameteroftheorbit; OrbEar: orbittoeardistance,
California Academy of Sciences, San Francisco; CUMZ fromtheanterioredgeoftheearopeningtotheposterioredge
R: Chulalongkorn University Museum of Zoology (Reptiles), of the orbit; RH: rostral height, the maximum height of the
Bangkok; IRSNB: Institut Royal des Sciences Naturelles rostralshield; RW:rostralwidth, thedistancebetweenborder
de Belgique, Brussels; KZM: Khorat Zoo Museum, Nakhon of rostral shield and the first supralabial scales on right and
Ratchasima; MNHN: Muséum National d’Histoire Naturelle, left sides; SnOrb: snout to orbit distance, from the tip of the
Paris; PSUZC-RT: Prince of Songkhla University Zoological snouttotheanteriormostmarginoftheorbit; SVL:snout-vent
Collection,ReptileSection,Songkhla;QSMI:QueenSaovabha length,takenfromthetipofsnouttothevent;TailL:taillength,
MemorialInstitute,ThaiRedCrossSociety,Bangkok;THNHM: takenfromtheventtothetipofthetail,originalorregenerated;
ThailandNaturalHistoryMuseum, NationalScienceMuseum, TailW: tail width, taken at the base of the tail immediately
Technopolis, Pathum Thani; ZMMU: Zoological Museum, posteriortothepostcloacalswelling;TibiaL:tibialength,taken
Moscow State University, Moscow; ZISP: Zoological Institute, on the ventral surface from the posterior surface of the knee
Saint Petersburg. Comparisons with congeneric species are while flexed 90◦ to the base of heel. Meristic characters
basedonliteraturedataandondirectexaminationofpreserved abbreviations: DorTub: number of longitudinal rows of dorsal
specimens(AppendixI). tubercles at midbody; EnlFemSc: enlarged femoral scales;
ZoologicalResearch39(3):202–219,2018 203
FemPi: femoral pits; FemPo: femoral pores; FemPreclPo: waxy exudates); PreclPo: precloacal pores (deeper than
number of femoral and precloacal pores in continuous series; pits, and with waxy exudates); SL: supralabial scales; SLF4:
IL: infralabial scales; InterorbSc: interorbital scales; ParaTub: subdigitallamellaebeneath4thfinger(basalanddistallamellae
numberofparavertebraltuberclesbetweenthelimbsinsertions, combined); SLT4: subdigital lamellae beneath 4th toe (basal
counted in a straight line immediately left of the vertebral and distal lamellae combined); Ven: number of ventral scale
column; PreclPi: precloacal pits (shallow depressions without rows.
Figure1GeneraldistributionofthegenusCyrtodactylusinLaosandsurroundingareas
FordetailedlocalityinformationseeAppendixII.IconcolorscorrespondtothespeciesgroupsasshowninFigure2:A:Yellow,C.interdigitalisspeciesgroup;
B:Green,C.pulchellusspeciesgroup;C:Red,C.wayakoneispeciesgroup;D:Blue,C.irregularisspeciesgroup;E:Purple,C.phongnhakebangensisspecies
group;dotiniconcenterindicatestypelocality;largedotindicatestypelocalitiesforseveralspeciesofCyrtodactylus.MYN—Myanmar.
DNAisolation,PCR,andsequencing measured in 1 µL using NanoDrop 2000 (Thermo Scientific,
USA),andconsequentlyadjustedtoca.100ngDNA/µL.
For molecular phylogenetic analyses, total genomic DNA ThemitochondrialcytochromeoxidasesubunitI(COI)was
was extracted from ethanol-preserved femoral muscle tissue selectedasageneticmarkertoclarifythetaxonomicposition
(Table1)usingstandardphenol-chloroform-proteinaseK(final of the newly discovered population of Cyrtodactylus. A
concentration1mg/mL)extractionprocedureswithconsequent total of 655 bp of COI was amplified using a mitochondrial
isopropanol precipitation (protocols followed Hillis et al., 1996 marker widely used as a barcoding marker for vertebrates,
and Sambrook et al., 1989). The isolated total genomic DNA including both reptiles and amphibians (Murphy et al.,
wasvisualizedusingagaroseelectrophoresisinthepresence 2013; Nagy et al., 2012; Smith et al., 2008), and
of ethidium bromide. The concentration of total DNA was
204 www.zoores.ac.cn
which proved to be useful for species identification and Ronquist & Huelsenbeck, 2003). Metropolis-coupled Markov
assessment of cryptic diversity in various groups of chainMonteCarlo(MCMCMC)analyseswererunwithonecold
lizards, including the genus Cyrtodactylus (Brennan et al., chain and three heated chains for four million generations and
2017; Hartmann et al., 2013; Luu et al., 2016a; Nazarov et sampledevery1000generations. FiveindependentMCMCMC
al., 2012, 2014; Nguyen et al., 2013, 2014, 2017; Solovyeva runswereperformedand1000treeswerediscardedasburn-in.
et al., 2011). Primers used both for PCR and sequencing Confidenceintreetopologywasassessedbyposteriorprobability
weretheVF1-d(5(cid:48)-TTCTCAACCAACCACAARGAYATYGG-3(cid:48)) (BI PP) (Huelsenbeck & Ronquist, 2001). The ML analyses
andtheVR1-d(5(cid:48)-TAGACTTCTGGGTGGCCRAARAAYCA-3’) were conducted using Treefinder (Jobb et al., 2004). The COI
(following Ivanova et al., 2006). The obtained fragments sequenceofCyrtodactyluselok Dringwasusedasanoutgroup
were sequenced in both directions for each sample, and a following Luu et al. (2016a). Confidence in tree topology was
consensus sequence was generated. PCR analyses were testedbynon-parametricbootstrapanalysis(MLBS)with1000
performed in 25 µL reactions using ca. 50 ng genomic DNA, replicates(Felsenstein,2004). Weapriori regardedtreenodes
10 pmol of each primer, 15 nmol of each dNTP, 50 nmol with bootstrap (ML BS) values of 70% or greater and posterior
additionalMgCl ,TaqPCRbuffer(10mmol/LTris-HCl,pH8.3, probabilities(BIPP)valuesover0.95assufficientlyresolved,ML
2
50mmol/LKCl,1.1mmol/LMgCl ,and0.01%gelatin),and1U BSvaluesbetween70%and50%(andBIPPbetween0.95and
2
ofTaqDNApolymerase.ThePCRconditionsfortheCOIgene 0.90) were regarded as tendencies, and ML BS values below
fragmentfollowedNazarovetal. (2012)andincludedaninitial 50%(BIPPbelow0.90)wereconsideredunresolved(Felsenstein,
denaturationstepat95◦Cfor3min; 5cyclesat95◦Cfor30 2004;Huelsenbeck&Hillis,1993).
s, annealingat45◦Cfor1min, extensionat72◦Cfor2min,
followedwith35cyclesat95◦Cfor30s,annealingat51◦Cfor RESULTS
1min,extensionat72◦Cfor2min,andafinalextensionat72
Geneticdifferentiation
◦Cfor5min.
Sequencedata
ThePCRproductswereloadedonto1.5%agarosegelsin
The final alignment of the examined mtDNA COI gene
thepresenceofethidiumbromideandvisualizedusingagarose
fragment consisted of 672 sites: 386 sites were conserved
electrophoresis. If distinct bands were produced, products
and 286 sites were variable, of which 270 were found to be
were purified using 2 µL, from a 1:4 dilution of ExoSapIt
potentially parsimony-informative. The transition-transversion
(Amersham, UK), per 5 µL of PCR product prior to cycle
bias(R)wasestimatedas3.66. Nucleotidefrequencieswere
sequencing. A 10 µL sequencing reaction included 2 µL of
A=23.14%,T=26.77%,C=30.53%,andG=19.95%.
template, 2.5 µL of sequencing buffer, 0.8 µL of 10 pmol
primer, 0.4 µL of BigDye Terminator version 3.1 Sequencing mtDNAgenealogy
Standard (Applied Biosystems), and 4.2 µL of water. The
Resultsofthephylogeneticanalysesimplementedinthepresent
cycle sequencing reaction included 35 cycles of 10 s at 96
paper are shown in Figure 2. The BI and ML analyses
◦C, 10 s at 50◦ C, and 4 min at 60 ◦C. Cycle sequencing
resulted in essentially similar topologies slightly differing from
products were purified by ethanol precipitation. Sequence
each other only in associations at several poorly supported
data collection and visualization were performed on an ABI
basal nodes. In general, the genetic differentiation of the
3730xlautomatedsequencer(AppliedBiosystems,USA).The
Cyrtodactylus phongnhakebangensis species group members
obtained sequences were deposited in GenBank under the
washighlyconsistentwiththeresultsreportedinpreviousstudies
accessionnumbersMG791873–MG791875(Table1).
by Nazarov et al. (2014) and Luu et al. (2016a). The
Phylogeneticanalyses partial fragment of the COI gene strongly supports monophyly
A total of 66 COI sequences of congeners were obtained oftheC.phongnhakebangensis(1.0/100,hereafternodesupport
from GenBank for phylogenetic analyses, including the three values are given for BI PP/ML BS, respectively), C. wayakonei
sequencesobtainedinthisstudy(Table1). (1.0/86),andC.irregularis (0.99/83)speciesgroups. Withinthe
Nucleotide sequences were initially aligned using ClustalX C. phongnhakebangensis species group, C. pageli Schneider,
1.81 (Thompson et al., 1997) with default parameters, and Nguyen, Schmitz, Kingsada, Auer & Ziegler, 2011 formed a
then optimized manually in BioEdit 7.0.5.2 (Hall, 1999) and sister lineage with respect to all other members of the C.
MEGA 6.0 (Tamura et al., 2013). Mean uncorrected genetic phongnhakebangensis species group (0.97/75). The remaining
distances(P-distances)betweensequencesweredetermined members of the C. phongnhakebangensis species group were
with MEGA 6.0. MODELTEST v.3.06 (Posada & Crandall, clustered in two subclades with moderate to strong support
1998) was used to estimate the optimal models of DNA values: (1)subcladeconsistingofC.khammouanensisNazarov,
evolution. The best-fitting model selected for COI dataset Poyarkov, Orlov, Nguyen, Milto, Martynov, Konstantinov &
was K80+I+G for the first and the second codon positions Chulisov,2014,C.rufford Luu,Calame,Nguyen,Le,Bonkowski
andGTR+Gforthethirdcodonposition, assuggestedbythe &Ziegler,2016,C.bansocensisLuu,Nguyen,Le,Bonkowski&
AkaikeInformationCriterion(AIC). Ziegler,2016,C.soudthichaki Luu,Calame,Nguyen,Bonkowski
Phylogenetictreeswereinferredusingtwodifferentmethods: & Ziegler, 2015, C. jaegeri Luu, Calame, Bonkowski, Nguyen
Bayesian inference (BI) and maximum likelihood (ML). BI was & Ziegler, 2014, C. sommerladi Luu, Bonkowski, Nguyen, Le,
conducted in MrBayes 3.1.2 (Huelsenbeck & Ronquist, 2001; Schneider,Ngo&Ziegler,2016,andC.roesleriZiegler,Nazarov,
ZoologicalResearch39(3):202–219,2018 205
Orlov, Nguyen, Vu, Dang, Dinh&Schmitz, 2010(0.95/64); and theBolikhamxayProvinceofLaos(1.0/100).
(2) subclade consisting of C. hinnamnoensis Luu, Bonkowski,
Nguyen, Le, Schneider, Ngo & Ziegler, 2016, C. darevskii Geneticdistances
Nazarov,Poyarkov,Orlov,Nguyen,Milto,Martynov,Konstantinov The uncorrected genetic P-distances among and within
& Chulisov, 2014, C. calamei Luu, Bonkowski, Nguyen, Le, the COI gene fragment of the studied members of
Schneider,Ngo&Ziegler,2016,C.phongnhakebangensisZiegler, the Cyrtodactylus phongnhakebangensis species group are
Rösler, Herrmann & Vu, 2002, C. lomyenensis Ngo & Pauwels, summarized in Table 2. The observed interspecific distances
2010, C. multiporus Nazarov, Poyarkov, Orlov, Nguyen, Milto, in the COI gene within the C. phongnhakebangensis species
Martynov, Konstantinov & Chulisov, 2014, C. teyniei David, group varied from P=4.2% (between C. darevskii and
Nguyen, Schneider & Ziegler, 2011, C. cf. jarujini Ulber,1993, C. hinnamnoensis) to P=19.4% (between C. soudthichaki
andCyrtodactylussp.fromBanThathom,XiangkhoangProvince, and C. hinnamnoensis) (Table 2). These values slightly
Laos(0.94/78). overlapped with interspecific comparisons between the C.
The newly discovered Cyrtodactylus population from Ban phongnhakebangensis species group members and the
Thathom, Xiangkhoang Province, was placed within the first outgroup Cyrtodactylus species (18.1%<P<24.4%, data not
subclade of the C. phongnhakebangensis species group and shown in Table 2). The observed intraspecific distances in
wasclusteredinonegroupwithC.multiporus,C.teyniei,and ouranalysisvariedfromP=0%toP=3.8%,withthelastvalue
C.cf.jarujini(1.0/99).Amongthesespecies,theBanThathom corresponding to the genetic differentiation between mtDNA
populationappearedtobeasisterlineageofC.cf.jarujinifrom lineagesofC.pageli (Table2).
Figure2Bayesianinference(BI)treefortheCyrtodactylusphongnhakebangensisspeciesgroupandotherCyrtodactylusspecies
groupsinhabitingLaosandadjacentareasbasedonanalysisofCOIgenefragments
CladecolorscorrespondtoiconcolorsinFigure1.ValuesatnodescorrespondtoBIPP/MLBSsupportvalues,respectively.SequenceofC.elokwasusedas
anoutgroup.InsetshowsC.roesleri,arepresentativeoftheC.phongnhakebangensisspeciesgroup.
206 www.zoores.ac.cn
Table1Specimensandsequencesof CyrtodactylusrepresentativesusedinmolecularanalysesofmtDNAgenefragment
No. Species GenBankaccessionNo. Locality VoucherID
1 Cyrtodactylusbadenensis KF929505 Vietnam,TayNinhProv. KIZ13689
2 C.bansocensis KU175573 Laos,KhammouaneProv. VNUF–R.2015.20
3 C.bansocensis KU175574 Laos,KhammouaneProv. NUOL-R-2015.21
4 C.bobrovi KT004368 Vietnam,HoaBinhProv. IEBR-A.2015.30
5 C.bobrovi KT004369 Vietnam,HoaBinhProv. VNMN-A.2015.61
6 C.calamei KX064043 Laos,KhammouaneProv. NUOL-R-2015.22
7 C.calamei KX064044 Laos,KhammouaneProv. VNUF-R.2015.28
8 C.cryptus KF169971 Vietnam,QuangBinhProv. PNKB3
9 C.cryptus KF169972 Vietnam,QuangBinhProv. PNKB4
10 C.cryptus KX064038 Laos,KhammouaneProv. VNUF-R.2014.69
11 C.elok HM888478 Malaysia ZMMU-RAN1991
12 C.darevskii HQ967221 Laos,KhammouaneProv. ZISP-FN256
13 C.darevskii HQ967223 Laos,KhammouaneProv. ZISP-FN223
14 C.hinnamnoensis KX064045 Laos,KhammouaneProv. IEBR-A.2013.89
15 C.hinnamnoensis KX064046 Laos,KhammouaneProv. IEBR-A.2013.90
16 C.hinnamnoensis KX064047 Laos,KhammouaneProv. VNUF-R.2015.11
17 C.hinnamnoensis KX064048 Laos,KhammouaneProv. VNUF-R.2015.3
18 C.hinnamnoensis KX064049 Laos,KhammouaneProv. NUOL-R-2015.9
19 C.lomyenensis KJ817436 Laos,KhammouaneProv. IEBR-KM2012.54
20 C.lomyenensis KP199942 Laos,KhammouaneProv. IEBR-KM2012.52
21 C.interdigitalis KX077901 Laos,KhammouaneProv. VNUF-R.2014.50
22 C.jaegeri KT004364 Laos,KhammouaneProv. IEBR-A.2013.55
23 C.jaegeri KT004365 Laos,KhammouaneProv. NUOL-R.2013.1
24 C.jaegeri KT004366 Laos,KhammouaneProv. VFU-TK914
25 C.cf.jarujini KX077907 Laos,BolikhamxayProv. VNUF-R.2015.7
26 C.khammouanensis HM888467 Laos,KhammouaneProv. ZISP-FN191
27 C.khammouanensis HM888469 Laos,KhammouaneProv. ZISP-FN257
28 C.kingsadai KF188432 Vietnam,PhuYenProv. IEBR-A.2013.3
29 C.cf.martini KF929537 China,YunnanProv. KIZ201103
30 C.multiporus HM888472 Laos,KhammouaneProv. ZISP-FN3
31 C.multiporus HM888471 Laos,KhammouaneProv. ZISP-FN2
32 C.otai KT004370 Vietnam,HoaBinhProv. IEBR-A.2015.26
33 C.otai KT004371 Vietnam,HoaBinhProv. IEBR-A.2015.27
34 C.puhuensis KF929529 Vietnam,ThanhHoaProv. KIZ11665
35 C.aff.pulchellus HQ967202 Malaysia ZMMU-R-12643-3
36 C.aff.pulchellus HQ967203 Malaysia ZMMU-R-12643-4
37 C.pageli KJ817431 Laos,VientianeProv. ZFMK91827
38 C.pageli KX077902 Laos,VientianeProv. NQT2010.36
39 C.pageli KX077903 Laos,VientianeProv. NQT2010.37
40 C.phongnhakebangensis KF929526 Vietnam,QuangBinhProv. PNKB2011.30
41 C.phongnhakebangensis KF929527 Vietnam,QuangBinhProv. PNKB2011.32
42 C.pseudoquadrivirgatus KF169963 Vietnam,ThuaThien-HueProv. ITBCZ3001
43 Cyrtodactylussp.1 KP199949 Vietnam,DaNang,BaNa ZMMU-R-13095-2
44 C.quadrivirgatus HM888465 Malaysia ZMMU-RAN1989
45 C.quadrivirgatus HM888466 Malaysia ZMMU-RAN1990
46 C.roesleri KF929532 Vietnam,QuangBinhProv. PNKB2011.34
47 C.roesleri KF929531 Vietnam,QuangBinhProv. PNKB2011.3
48 C.rufford KU175572 Laos,KhammouaneProv. VFU-R.2015.14
49 C.sommerladi KX064039 Laos,KhammouaneProv. IEBR-A.2015.37
50 C.sommerladi KX064040 Laos,KhammouaneProv. VNUF-R.2013.22
51 C.sommerladi KX064041 Laos,KhammouaneProv. VNUF-R.2013.87
52 C.sommerladi KX064042 Laos,KhammouaneProv. IEBR-A.2015.39
53 C.spelaeus KP199947 Laos,VientianeProv. ZMMU-R-13980-3
54 C.spelaeus KP199948 Laos,VientianeProv. ZMMU-R-13980-1
55 C.soudthichaki KX077904 Laos,KhammouaneProv. NUOL-R-2015.5
56 C.soudthichaki KX077905 Laos,KhammouaneProv. VFU-R.2015.18
57 C.soudthichaki KX077906 Laos,KhammouaneProv. IEBR-A.2015.34
58 C.teyniei KJ817430 Laos,KhammouaneProv. IEBR-KM2012.77
59 C.teyniei KP199945 Laos,KhammouaneProv. IEBR-KM2012.77
60 C.vilaphongi KJ817434 Laos,LuangPrabangProv. NUOL-R-2013.5
61 C.vilaphongi KJ817435 Laos,LuangPrabangProv. IEBR-A.2013.103
62 C.wayakonei KJ817438 Laos,LuangNamThaProv. ZFMK91016
63 C.wayakonei KP199950 Laos,LuangNamThaProv. ZMMU-R-13981-1
64 Cyrtodactylusthathomensissp.nov. MG791873 Laos,XiangkhouangProv. ZMMU-R-14919-1
65 Cyrtodactylusthathomensissp.nov. MG791874 Laos,XiangkhouangProv. ZISP29731
66 Cyrtodactylusthathomensissp.nov. MG791875 Laos,XiangkhouangProv. ZMMUR-15384
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Table 2 Uncorrected P-distances (percentages) between COI sequences of the Cyrtodactylus phongnhakebangensis species
groupmembersincludedinphylogeneticanalyses(belowthediagonal),andstandarderrorestimates(abovethediagonal)
No. Species 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17
1 C.thathomensis sp.nov. 0.0 1.8 1.7 1.7 1.7 1.8 1.5 1.5 1.4 1.5 1.8 1.7 1.4 1.0 1.3 1.2 1.8
2 C.khammouanensis 18.3 0.2 1.4 1.4 1.4 1.5 1.4 1.3 1.4 1.5 1.3 1.4 1.4 1.8 1.6 1.5 1.6
3 C.rufford 18.9 11.5 — 1.3 1.6 1.5 1.6 1.5 1.6 1.5 1.5 1.5 1.5 1.8 1.4 1.5 1.7
4 C.bansocensis 17.1 12.1 11.2 0.2 1.6 1.5 1.4 1.3 1.7 1.8 1.5 1.6 1.4 1.6 1.4 1.6 1.6
5 C.soudthichaki 18.4 13.3 14.5 12.2 0.1 1.6 1.3 1.4 1.8 1.8 1.6 1.7 1.8 1.6 1.5 1.4 1.4
6 C.jaegeri 18.5 14.6 14.5 14.8 13.5 0.1 1.6 1.5 1.9 1.8 1.8 1.7 1.4 1.8 1.7 1.8 1.7
7 C.sommerladi 17.7 16.7 17.9 16.0 15.5 16.3 0.1 0.8 1.5 1.6 1.4 1.4 1.6 1.5 1.5 1.7 1.4
8 C.roesleri 17.9 16.5 16.4 15.9 15.2 15.4 6.0 0.2 1.6 1.7 1.4 1.4 1.5 1.4 1.4 1.6 1.3
9 C.hinnamnoensis 16.4 16.6 18.7 17.5 19.4 18.1 16.3 17.1 0.9 0.8 0.9 1.2 1.5 1.4 1.5 1.4 1.6
10 C.darevskii 16.2 14.8 18.5 16.7 18.5 17.8 15.7 17.3 4.2 0.0 1.0 1.4 1.6 1.6 1.8 1.6 1.6
11 C.calamei 17.3 13.7 15.6 16.4 17.6 17.3 15.2 15.5 5.2 5.6 0.0 1.1 1.4 1.8 1.6 1.6 1.5
12 C.phongnhakebangensis 16.0 15.5 17.6 15.4 17.7 16.7 16.3 15.3 9.1 9.6 7.8 0.0 1.3 1.6 1.7 1.8 1.4
13 C.lomyenensis 15.5 15.6 17.4 16.0 18.8 17.1 17.8 17.5 15.4 14.5 15.0 14.6 0.2 1.4 1.4 1.3 1.6
14 C.cf.jarujini 4.5 16.1 17.3 18.0 19.0 18.8 18.1 17.4 16.5 16.7 17.1 17.1 15.2 — 1.2 1.1 1.6
15 C.multiporus 9.5 16.6 15.8 16.4 18.3 16.1 17.0 17.0 15.4 15.6 15.5 15.3 14.4 9.3 0.0 1.0 1.5
16 C.teyniei 9.3 17.2 17.3 18.3 17.4 18.1 17.7 17.5 15.3 15.6 14.2 15.3 14.5 9.3 6.0 0.0 1.5
17 C.pageli 18.5 18.3 17.8 18.8 18.8 18.0 16.2 16.6 18.8 18.2 16.9 17.8 17.7 18.0 16.5 17.5 3.8
IngroupmeanuncorrectedP-distancesareshownonthediagonal.
The newly discovered Cyrtodactylus population from Ban distinguishedfromallothercongenericspeciesbyitsmedium
Thathom,XiangkhoangProvince,Laos,isgeneticallydivergent bodysize(maximalknownSVL75.5mm); dorsaltuberclesin
from all other members of the C. phongnhakebangensis 14–18 rows at midbody; midbody scale rows 30–36 across
species group, and it is most closely related to C. multiporus belly between ventrolateral skin folds; a continuous series
(P=9.5%), C. teyniei (P=9.3%), and C. cf. jarujini (P=4.5%). of 10–12 pore-bearing (male) or pitted (females), enlarged
The last population from Bolikhamxay Province of Laos was precloacal scales, separated by a diastema from a series of
tentatively identified as C. cf. jarujini by Luu et al. (2016a); enlargedfemoralscalesbearing18or19pores(male)or1–9
however,itstaxonomicstatusrequiresfurtherconfirmation. pits (females) on each femur; absence of precloacal groove;
transverselyenlargedmediansubcaudalscales;andfourdark
Systematics
dorsalbandsbetweenlimbinsertions.
Based on the morphological, chromatical, and genetic
distinctiveness of the newly discovered Cyrtodactylus Description of holotype: Adult male. SVL 70.3 mm.
population from all other populations in northern Laos and TailL 79.0 mm (last 54 mm regenerated). Head relatively
neighboring areas (see Diagnosis and Comparisons), and long (HeadL 20.1; HeadL/SVL 0.29) and wide (HeadW 13.4;
alsoontheresultsofphylogeneticanalysesofthepartialCOI HeadW/HeadL 0.67), not markedly depressed (HeadH 8.4),
gene fragment, indicating that this population represents a distinct from neck. Loreal region inflated, canthus rostralis
clearly distinct mtDNA lineage, different from all congeners slightly prominent. Snout elongate (SnOrb/HeadL 0.41),
for which homologous sequences are available (see Results), rounded,longerthanorbitdiameter(OrbD/SnOrb0.62).Scales
we conclude that the Ban Thathom population represents a on snout small, rounded to oval, granular to weakly conical,
previously undescribed new species, which we describe and mostlyhomogeneous,largerthanthoseoncrown,interorbital,
nameherein. andoccipitalregions. Eyerelativelylarge(OrbD/HeadL0.25);
pupil vertical with crenelated margins; supraciliaries short,
Cyrtodactylusthathomensissp.nov.(Figures3–6) larger anteriorly. Ear opening vertically oval, of moderate
size (EarL/HeadL 0.06); orbit to ear distance subequal to
Holotype: ZMMU R-14919-1 (Figure 3); an adult male from
orbit diameter (OrbEar/OrbD 1.02). Rostral much wider (3.5
the north-western slope of a limestone hill (N18◦59(cid:48)48.9(cid:48)(cid:48),
mm) than deep (2.0 mm); rostral crease straight, starting
E103◦35(cid:48)30.6(cid:48)(cid:48);alt.271ma.s.l.)nearBan(=Village)Thathom,
from the upper middle of the rostral, going down half the
XiangkhoangProvince,Laos. Collectedon14May2008byE.
rostral height. Two enlarged supranasals separated from
L.KonstantinovandA.S.Chulisov.
one another by one internasal. Rostral contacting first
supralabial on each side, nostrils, two supranasals and one
Paratypes: ZMMU R-15384 and ZISP 29731; two adult
internasal. Nostrils rounded, more or less laterally directed,
femalesfromthesamelocality
each surrounded by supranasal, rostral, first supralabial and
Diagnosis: Cyrtodactylus thathomensis sp. nov. can be two postnasals. Three or four rows of small scales separate
208 www.zoores.ac.cn
orbitfromsupralabials. Mentaltriangular,wider(3.4mm)than (in total three granules contact the postmentals) (Figure 3E).
deep(2.2mm). Asinglepairofgreatlyenlargedpostmentals Supralabials to mid-orbital position 9/9, enlarged supralabials
inbroadcontactbehindmental,eachborderedanteromedially toangleofjaws10/11. Infralabials9/9(Figure3D).Interorbital
bymental,anterolaterallybyfirstinfralabial,posterolaterallyby scalerowsacrossnarrowestpointoffrontalbone30.
anenlargedlateralchinshield,andposteriorlybytwogranules
Figure3MaleholotypeofCyrtodactylusthathomensissp.nov.(ZMMUR-14919)
A:Generalviewofholotypeinsitu;B:Precloacalregionwithrowoffemoralandprecloacalporesseparatedbydiastema;C:Postcloacalspurs(seearrow);D:
Lateralsideoftheheadwithsupralabialsandinfralabialsscalation;E:Ventralsurfaceoftheheadwithmentalscalation;F:Dorsumsurfacewiththerowsof
enlargedtubercles.PhotosbyE.L.Konstantinov(A)andR.A.Nazarov(B–F).
ZoologicalResearch39(3):202–219,2018 209
Body moderately slender, relatively short (AG/SVL 0.43) Variation: Morphometric and meristic values for the type
with poorly defined, non-denticulate, ventrolateral skin folds. series are provided in Table 3. Morphological and coloration
Dorsal scales weakly heterogeneous, domed, with tubercles characters of the paratypes agree in most respects with
aboutfourtimessizeofadjacentdorsalscalesextendingfrom those of the holotype, differing only in minor details (Table
neck onto tail base, smaller tubercles on postocular region, 3 and remarks hereafter). Rostral crease in paratypes like
crown and occiput; tubercles smooth or bearing a very small in holotype. One internasal in paratype ZISP 29731 like in
keel, tubercles on posterior trunk and sacral region most holotype(Figure4C),nointernasalinparatypeZMMUR-15384
prominent. Dorsal tubercles in 18 rows at midbody, typically (hence supranasals in large contact) (Figure 5C). The female
separatedfromoneanotherbytwodorsalgranules(Figure3F). paratypeZISP29731showsacontinuousrowof52enlarged
Paravertebral tubercles 26. Ventral scales larger than dorsal femoral-precloacalscalesincluding,fromlefttoright: 1pitless
scales, smooth, oval and subimbricate, largest in precloacal scale + 9 scales with pits + 12 pitless scales + 10 precloacal
region. Midbodyscalerowsacrossbellybetweenventrolateral scales with pits + 11 pitless scales + 9 scales with pits + 1
folds30. Gularregionwithhomogeneous,smooth,juxtaposed pitlessscale(Figure4E).ThefemaleparatypeZMMUR-15384
granularscales. Apatchofenlargedprecloacalscalesontop shows a continuous row of 55 enlarged femoral-precloacal
of which lies a continuous series of ten pore-bearing scales, scales including, from left to right: 10 scales with pits + 11
separated by a diastema of 3/4 enlarged poreless scales pitless scales + 12 enlarged precloacal scales with pits + 12
fromacontinuousseriesof19/18pore-bearingfemoralscales. pitlessscales+8scaleswithpits+2pitlessscales(Figure5E).
Enlargedfemoralscalesnearlytwotimesthesizeofthescales General background dorsal color in preservative lighter in the
of the adjacent anterior scale row (Figure 3B). No precloacal female paratypes than in the male holotype. The paratypes’
groove.Hemipenealbulgesevident.Postcloacalspursbearing tails, original and complete, show nine dark brown bands,
6/5enlargedconicalscales(Figure3C). andtheyarelongerthanSVL(TailL/SVLratio1.16–1.27; see
Scales on palm and sole smooth, rounded to oval or Figures4A–5A,Figure6).
hexagonal, slightly domed. Scalation on dorsal surfaces of
Comparison with congeneric species: Synoptic tables
hind limbs similar to body dorsum with enlarged tubercles
comparing the main morphological characters of Lao
interspersed among smaller scales; tubercles smaller and
Cyrtodactylus with the species known from adjacent regions
rare on forelimbs. Forelimbs and hind limbs moderately
were provided by Teynié & David (2014: 471–472) and Luu
long(ForeaL/SVL0.16, TibiaL/SVL0.19), moderatelyslender.
et al. (2016a, 2016b, 2016c). Among these species, we
Digits long, slender, inflected at interphalangeal joints, all
are comparing hereafter Cyrtodactylus thathomensis sp. nov.
bearing robust, slightly recurved claws. Basal subdigital
with all congeneric species found within a 500-km radius from
lamellaebroad,ovaltorectangular,withoutscansorialsurfaces;
its type-locality (a radius far superior to any maximal distance
lamellae distal to digital inflection narrow; SLF4 17/17; SLT4
betweentwolocalitiesknownforanyspeciesintheIndochinese
20/20. Subcaudals scales larger than supracaudal scales,
Region, Thailand and Myanmar, cf. maps provided by Ellis &
formingarowofstronglyenlargedtransverseplates.
Pauwels, 2012; Nazarov et al., 2014: Figure 1; Grismer et al.,
Coloration in preservative: Dorsal ground color of head, 2015:Figure1–consideringthatCyrtodactylusintermediusisa
neck,body,limbsandtaillightbrown. Dorsalsurfaceofhead speciescomplexcomposedofatleastsixspecies,loc. cit.: 114;
with irregular dark brown markings. Rostral, supralabials and Nguyenetal.,2014,2017:Figure1).
infralabialsdarkbrown,posterioronesheavilymaculatedwith By its possession of transversely enlarged subcaudals,
beige. Oneachsideapostocularstripereachingthenapebut the new species is readily distinguished from Cyrtodactylus
notmeetingtheoneoftheoppositesideasitbreaksintospots angularis ( Smith, 1921), C. buchardi David, Teynié & Ohler,
(i.e., a discontinuous nuchal collar). Upper surface of limbs 2004, C. cryptus Heidrich, Rösler, Vu, Böhme & Ziegler,
showing irregular dark brown bands. Dorsum showing four 2007, C. jarujini, C. papilionoides Ulber & Grossmann, 1991,
dark brown bands between limb insertions. Each of the four C. pseudoquadrivirgatus Rösler, Nguyen, Vu, Ngo & Ziegler,
bandsondorsumposteriorlylimitedbyadiscontinuousseries 2008, and C. vilaphongi Schneider, Nguyen, Le, Nophaseud,
ofwhitishtubercles,similarlytothebandaboveshoulders,the Bonkowski & Ziegler, 2014. The possession of enlarged
one on the neck and the one above sacrum. Original part of femoralscalesseparatesCyrtodactylusthathomensissp. nov.
thetailshowingthreedarkbrownbands. Regeneratedpartof from C. bobrovi Nguyen, Le, Pham, Ngo, Hoang, Pham &
thetaillightbrown. Undersurfacesofthehead, throat, venter Ziegler, 2015, C. buchardi, C. chauquangensis Hoang, Orlov,
andmembersuniformlybeige.Colorationoftheholotypeinlife Ananjeva, Johns, Hoang & Dau, 2007, C. cryptus, C. otai
is shown in Figure 3A. In life its dorsal ground color is darker Nguyen, Le, Pham, Ngo, Hoang, Pham & Ziegler, 2015,
than in preservative. The tubercles posteriorly bordering the C. pageli, C. pseudoquadrivirgatus, C. spelaeus Nazarov,
four bands on dorsum are lighter and more contrasting than Poyarkov, Orlov, Nguyen, Milto, Martynov, Konstantinov &
afterpreservation. Theyellowcoloroftheouterextremitiesof Chulisov, 2014, C. vilaphongi and C. wayakonei Nguyen,
thesupraciliariesdisappearsinpreservative. Kingsada,Rösler,Auer&Ziegler,2010.
210 www.zoores.ac.cn
Figure4FemaleparatypeofCyrtodactylusthathomensissp.nov.(ZISP29731)
A:Generalviewofpreservedparatypewithoriginaltail;B:Lateralsideoftheheadwithsupralabialsandinfralabialsscalation;C:Shapeofrostralandsurrounding
scales;D:Ventralsurfaceoftheheadwithmentalscalation;E:Precloacalregionwithrowofenlargedfemoralandprecloacalscaleswithpitandpitlessscales;
F:Medialrowofenlargedsubcaudalscales.PhotosbyR.A.Nazarov.
ZoologicalResearch39(3):202–219,2018 211