Труды Зоологического института РАН
Том 318, № 4, 2014, c. 391–423
УДК 575+598.1
A REVIEW OF GENUS CYRTODACTYLUS (REPTILIA: SAURIA: GEKKONIDAE)
IN FAUNA OF LAOS WITH DESCRIPTION OF FOUR NEW SPECIES
R.A. Nazarov1*, N.A. Poyarkov Jr.2,3, N.L. Orlov4, N.S. Nguyen5, K.D. Milto4, A.A. Martynov2,
E.L. Konstantinov 6 and A.S. Chulisov 6
1
Zoological Museum, Moscow State University, B. Nikitskaya St. 6, 125009 Moscow, Russia; e-mail: r_nazarov@mail.ru;
2
Department of Vertebrate Zoology, Biological Faculty, Lomonosov Moscow State University, Leninskiye Gory, 119991
Moscow, GSP-1, Russia;.e-mail: n.poyarkov@gmail.com
3
Joint Russian-Vietnamese Tropical Research and Technological Center under the A.N. Severtsov Institute of Ecology
and Evolution RAS, South Branch, 3, Street 3/2, 10 District, Ho Chi Minh City, Vietnam
4
Zoological Institute of the Russian Academy of Sciences, Universitetskaya Emb. 199034 Saint-Petersburg, Russia;
e-mail: azemiops@zin.ru
5
Institute of Tropical Biology, Vietnam Academy of Science and Technology (VAST), 85 Tran Quoc Toan St., District 3,
Ho Chi Minh City, Vietnam; e-mail: ngocsangitb@yahoo.com
6
Institute of Natural Sciences, Kaluga State University named after K.I. Tsiolkovskii, Stepana Razina St. 26, 248023 Kaluga,
Russia; e-mail: nepentes@list.ru
ABSTRACT
We describe four new species of the genus Cyrtodactylus based on the results of phylogenetic analysis of 673 bp of
the COI mtDNA gene and morphological description of voucher specimens from Khammouane and Vientiane provinces, Laos. The three of the new species, namely Cyrtodactylus khammouanensis sp. nov., Cyrtodactylus darevskii sp.
nov. and Cyrtodactylus multiporus sp. nov., are described from a small area in monsoon tropical forests on limestone
in the environs of Nahome Village, Boulapha District, Khammouane Province, Central Annamites. Cyrtodactylus
darevskii sp. nov. differs from all other Laotian Cyrtodactylus by a combination of the following morphological
characters: (1) maximum SVL up to 100 mm; (2) dorsal pattern formed by the narrow dark nuchal band and 4–5
dark transverse breaking bands with light yellowish posterior edges between limbs; (4) 38–46 ventral scales; (5)
continuous series of 38–44 precloacal and femoral pores in males and 24–34 precloacal and femoral pores in females;
(6) preclocal groove lacking; (7) 4–5 postcloacal spurs present in both sexes; (8) one median row of subcaudal
scales. Cyrtodactylus khammouanensis sp. nov. appears to be closely related to the recently described C. jaegeri Luu
et. al., 2014 and can be distinguished from this species and the remaining Laotian congeners by a combination of the
following morphological features: (1) medium size, with a maximum SVL up to 73 mm; (2) males with continuous
series of 40–44 precloacal and femoral pores; (3) preclocal groove lacking; (4) 32–38 longitudinal rows of ventral
scales at midbody; (5) weakly keeled tubercles present on the dorsum and lacking on the head; (6) 5–6 enlarged
postcloacal spurs in both sexes; (8) one median row of subcaudal scales; (7) dorsal pattern consisting of wide dark
nuchal band and 4–5 dark wide transverse bands. Cyrtodactylus multiporus sp. nov. is distinguished by a combination
of the following morphological attributes: (1) maximum SVL of up to 98 mm; (2) dorsal patterns consisting of dark
irregular separate spots between limbs and on the dorsal surface of the head; (3) nuchal band absent; (4) 30–38
ventrals; (5) continuous series of 58–60 precloacal and femoral pores in males; (6) preclocal groove lacking; (7) 5–6
postcloacal spurs present in both sexes; (8) one median row of subcaudal scales. Cyrtodactylus spelaeus sp. nov. is
described from a cave in north of Vientiane Province of Laos and differs from other congeners by a combination of
the following characters: (1) maximum SVL of 98 mm; (2) dorsal pattern formed by oblong dark irregular butterflyshaped blotches with light contrast margins; (3) nuchal band with light edging consists of two separate parts, which
contact in occipital region; (4) 37–39 ventral scales; (5) 7–8 precloacal pores in males; (5) preclocal groove lacking;
(6) 2–3 postcloacal spurs present in both sexes; (7) ten rows of enlarged dorsal tubercles. Our study increase the
number of Cyrtodactylus species known for Laos to fifteen. COI DNA-barcoding is successfully applied to reveal
cryptic diversity within the genus Cyrtodactylus.
Key words: Central Annamites, COI barcoding, Cyrtodactylus darevskii sp. nov., Cyrtodactylus khammouanensis sp.
nov., Cyrtodactylus multiporus sp. nov., Cyrtodactylus spelaeus sp. nov., Laos, sympatry, taxonomy
*Corresponding author / Автор-корреспондент
392
R.A. Nazarov et al.
ОБЗОР РОДА CYRTODACTYLUS (REPTILIA: SAURIA: GEKKONIDAE)
ФАУНЫ ЛАОСА С ОПИСАНИЕМ ЧЕТЫРЕХ НОВЫХ ВИДОВ
Р.А. Назаров1, Н.А. Поярков2,3, Н.Л. Орлов4, Н.Ш. Нгуен5, К.Д. Мильто4, А.А. Мартынов2,
Е.Л. Константинов6 и А.С. Чулисов6
Научно-исследовательский Зоологический музей Московского государственного университета им. М.В. Ломоносова,
Б. Никитская ул. 6, Москва 125009, Россия; e-mail: r_nazarov@mail.ru
1
Биологический факультет Московского государственного университета им. М.В. Ломоносова, Ленинские Горы,
119991 Москва, ГСП 1,Россия; e-mail: n.poyarkov@gmail.com
2
Совместный Российско-Вьетнамский Научно-Исследовательский и Технологический Центр, Южное Отделение,
Хошимин, Вьетнам
3
4
Зоологический Институт Российской академии наук, Университетская наб. 1, 199034 Санкт-Петербург, Россия;
е-mail: azemiops@zin.ru
5
Институт Тропической Биологии, Вьетнамская Академия Наук и Технологии (ВАНТ), ул. Тян Куок Тоан 85, 3 район,
Хошимин, Вьетнам; e-mail: ngocsangitb@yahoo.com
6
Калужский государственный педагогический университет имени К.Э. Циолковского, ул. Степана Разина 26, 248023,
Калуга, Россия; e-mail: nepentes@list.ru
РЕЗЮМЕ
В статье по результатам филогенетического анализа фрагмента гена первой субъединицы цитохром
с-оксидахы мтДНК (COI) длиной 673 п.о. и морфологического исследования ваучерных экземпляров приводится описание четырех новых видов кривопалых гекконов рода Cyrtodactylus из провинций Кхаммуан и
Вьентьян, Лаос. Три из описанных новых для науки видов, а именно Cyrtodactylus khammouanensis sp. nov.,
Cyrtodactylus darevskii sp. nov. и Cyrtodactylus multiporus sp. nov., обнаружены на небольшой территории, покрытой мусонными тропическими лесами и карстовыми массивами, в окрестностях деревни Нахом, район
Буалапха, провинция Кхаммуан, центральная часть Аннамского хребта. Cyrtodactylus darevskii sp. nov. отличается от всех прочих кривопалых гекконов фауны следующей комбинацией морфологических признаков:
(1) максимальная длина тела SVL до 100 мм; (2) окраска дорзальной стороны тела сорфмирована узкой
темной затылочной (нухальной) полосой и 4–5 темными поперечными прерывистыми полосами с тонкой
желтой окантовкой между передними и задними конечностями; (4) 38–46 рядов брюшных чешуй; (5) непрерывающиеся ряды из 38–44 предклоакальных и бедренных пор у самцов и 24–34 предклоакальных и
бедренных пор у самок; (6) предклоакальная бороздка отсутствует; (7) 4–5 постклоакальных шипов развиты
обоих полов; (8) выражен один медиальный ряд подхвостовых чешуй. Cyrtodactylus khammouanensis sp. nov.
по-видимому близок к описанному недавно виду C. jaegeri Luu et. al., 2014, однако отличается от него вида,
как и других лаосских Cyrtodactylus, cледующими признаками внешней морфологии: (1) средний размер,
с максимальной длиной тела SVL до 73 мм; (2) самцы с непрерывным рядом из 40–44 предклоакальных и
бедренных пор; (3) предклоакальная бороздка отсутствует; (4) 32–38 продольных рядов брюшных чешуй
в средней части туловища; (5) слабо-килеватые бугорки расположены на дорзальной поверхности тела, но
отсутствуют на голове; (6) 5–6 увеличенных постклоакальных шипов как у самцов, так и у самок; (8) выражен один медиальный ряд подхвостовых чешуй; (7) рисунок спинной поверхности тела образован широкой
затылочной полосой и 4–5 темными поперечными полосами. Cyrtodactylus multiporus sp. nov. отличается от
других видов рода следующим сочетанием морфологических характеристик: (1) максимальная длина туловища SVL до 98 mm; (2) рисунок дорзальной поверхности тела представлена отдальными темными пятнами
неправильной формы, разбросанными по спине и заходящими на дорзальную поверхность головы; (3) затылочная полоса отсутствует; (4) 30–38 брюшных чешуй; (5) у самцов 58–60 преклоакальных и бедренных пор
расположенных в непрерывном ряду; (6) предклоакальная бороздка отсутствует; (7) 5–6 увеличенных постклоакальных шипов как у самцов, так и у самок; (8) выражен один медиальный ряд подхвостовых чешуй.
Cyrtodactylus spelaeus sp. nov. описан из пещеры в северной части провинции Вьентьян, Лаос, и отличается от
других кривопалых гекконов региона следующими морфологическими признаками: (1) максимальная длина тела SVL до 98 мм; (2) окраска спинной поверхности тела образована продолговатыми темными пятнами
со светлой окантовкой по краю; (3) затылочная полоса имеет тонкую светлую окантовку и состоит из двух
частей, конатктирующих в затылочной части черепа; (4) 37–39 брюшных чешуй; (5) 7–8 предклоакальных
A review of the genus Cyrtodactylus
393
пор у самцов; (5) предклоакальная бороздка отсутсвует; (6) 2–3 увеличенных постклоакальных шипа как
у самцов, так и у самок; (7) десять рядов увеличенных спинных бугорков. Наше исследование увеличивает
число видов рода Cyrtodactylus, известных для Лаоса, до пятнадцати. Мы также успешно применили методы
ДНК индентификации (COI ДНК-баркодинг) к оценке криптического разнообразия рода Cyrtodactylus.
Ключевые слова: центральный Аннам, COI barcoding, Cyrtodactylus darevskii sp. nov., Cyrtodactylus khammouanensis sp. nov., Cyrtodactylus multiporus sp. nov., Cyrtodactylus spelaeus sp. nov., Лаос, симпатрия, систематика
INTRODUCTION
The genus Cyrtodactylus is one of the largest genera of the gekkonid lizards and to date includes about
196 recognized species, distributed in the mainland
Southern and Southeast Asia, Himalayas, Indonesia,
northern Australia and Oceania (Uetz et al. 2014).
Cyrtodactylus thus forms the most diverse genus of
Gekkonidae to date, with Indochina representing
one of the centers of undiscovered species diversity
of this genus. Over the last decade, the diversity of
the genus has increased at several times as a result
of numerous descriptions of new species from poorly
studied areas of Indochina and Indonesia. In 1997
only 18 species of the genus Cyrtodactylus were
recorded from the huge territory of Southeast Asia
(Manthey and Grossmann 1997). The number of
described species has increased rapidly since 2007;
in Vietnam, for example, the number of recognized
species has increased from just 5 species known in
2006 to 33 species recognized in 2014 (Nguyen et al.
2014). There has been an enormous increase in the
number of new species of the genus from Southeast
Asia and Oceania, resulting in a total of at least 90
species recognized by 2008 (Ngo and Bauer 2008). At
least 44 species of the genus Cyrtodactylus were reported from Indochina – Thai region for 2011 (Uetz
et al. 2011).
The numerous new records and discoveries from
mainland Southeast Asia have been documented
mainly for Malaysia, Vietnam and Thailand. From
Laos only ten species of Cyrtodactylus have been
recorded to date: C. buchardi David, Teynié et Ohler,
2004, C. interdigitalis Ulber, 1993, Cyrtodactylus
jaegeri Luu, Calame, Bonkowski, Nguyen et Ziegler,
2014; C. jarujini Ulber, 1993, C. lomyenensis Ngo et
Pauwels, 2010, Cyrtodactylus pageli Schneider, Nguyen, Schmitz, Kingsada, Auer et Ziegler, 2011; Cyrtodactylus roesleri Ziegler, Nazarov, Orlov, Nguyen,
Vu, Dang, Dinh et Schmitz, 2010; C. teyniei David,
Nguyen, Schneider et Ziegler, 2011; Cyrtodactylus
vilaphongi Schneider, Nguyen, Duc Le, Nophaseud,
Bonkowski et Ziegler, 2014; C. wayakonei Nguyen,
Kingsada, Rösler, Auer et Ziegler, 2010; suggesting
that our knowledge of Laotian herpetofauna is still
far from being complete (Stuart 1999; David et al.
2004; Teynié et al. 2004; Teynié and David 2010; Ngo
and Pauwels 2010; Nguyen et al. 2010; David et al.
2011; Luu et al. 2013). Our field research in Laos at
2009 and 2011 led to discovery of four new populations of Cyrtodactylus of unclear taxonomic status.
Methods of DNA identification, or DNA barcoding, serve as important tools for biodiversity
assessment since they can be especially useful for
identification of cryptic species (Hebert et al. 2003,
2004; Smith et al. 2008). Herein we use DNA barcoding with COI to reexamine the taxonomic status of
known species of Laotian bent-toed geckos as well as
to explore the status of newly collected specimens in
the region. We use this gene, in part, because of its application in documenting genetic diversity through
the Cold Code project (Murphy et al. 2013), and also
referring to its successful application to biodiversity
assessment of the genus Cyrtodactylus (Nazarov et
al. 2012; Nguyen et al. 2013, 2014; Schneider et al.
2014a, 2014b). Our investigation results in the description of four new species of Cyrtodactylus based
on both genetics and morphology.
MATERIAL AND METHODS
Sample collection. Field work was conducted in
the Laotian provinces of Vientiane, Luang Prabang
and Khammouane in 2009–2013. Specimens were
anaesthetized, ethanol-fixed and subsequently deposited in the collections of the Zoological Institute,
St. Petersburg (ZIN), Russian Academy of Sciences,
Russia; Zoological Museum, Lomonosov Moscow
State University (ZMMU), Moscow, Russia. The
geographic position of the surveyed localities and
394
the distribution of the known Cyrtodactylus species
in the Central Annamite Mountains (Truong Son)
and adjacent regions of southern Indochina (central
and northern Vietnam, Laos, north-eastern part of
Thailand) are shown in Fig. 1. For localities of Cyrtodactylus distribution in Laos and adjacent areas
see Fig. 1.
Morphology. For the morphological descriptions
and comparisons, the following measurements (taken
with caliper rule to the nearest 0.1 mm) and scalation characters were used (after Darevsky, Szczerbak
1997; Bauer 2002, 2003; Bauer et al. 2002, 2003;
Ziegler et al. 2002; David et al. 2004; Nguyen et al.
2006; Hoang et al. 2007; Nazarov et al. 2008): Snoutvent length (SVL, from tip of snout to vent); tail
length (TailL, from vent to tip of tail); head length
(HeadL, distance between retroarticular process of
jaw and snout-tip); head width (HeadW, maximum
width of head); head height (HeadH, maximum
height of head, from occiput to underside of jaws);
orbital diameter (OrbD, greatest diameter of orbit);
snout to eye distance (SnEye, distance between
anterior most point of eye and tip of snout); ear
length (EarL, longest dimension of ear); trunk length
(TrunkL, distance from axilla to groin measured from
posterior edge of forelimb insertion to anterior edge
of hind limb insertion); shoulder length (LS); forearm length (ForeaL, from base of palm to elbow); femur length (FemurL); crus length (Crus L, from base
of heel to knee); length of finger IV (LF4); length of
toe IV (LT4); eye to ear distance (EyeEar, distance
from anterior edge of ear opening to posterior corner
of eye); scales across the belly in the middle of the
body (V); number of scales along the midbody from
mental shield to anterior edge of cloaca (SLB), supralabials (SL); infralabials (IL); precloacal pores
(PP); femoral pores (FP); enlarged femoral scales
(EFS); number of basal and distal subdigital lamellae
under fourth finger (SDL4A); number of basal and
distal subdigital lamellae under fourth toe (SDL4P),
number of longitudinal rows of enlarged tubercles
in the middle of dorsum between ventrolateral folds
(TubL); number of transversal rows of enlarged
tubercles between occipital region and middle of
sacrum along the middle of dorsum (TubW); number
of scales along middle of head, between occiput sinus
and supranasals (SLH); scales across head, between
top of ear openings (SAH); rostral width, between
supralabials (RW); rostral height, between anterior
and posterior margins of rostral (RH).
R.A. Nazarov et al.
We also obtained comparative morphological data
on Cyrtodactylus species found in Laos and surrounding parts of Vietnam and Thailand: C. bichnganae,
C. buchardi, C. chauquangensis, C. cryptus, C. cucphuongensis, C. huongsonensis, C. interdigitalis, C. jaegeri,
C. jarujini, C. kunyai, C. lomyenensis, C. martini,
C. pageli, C. phongnhakebangensis, C. pseudoquadrivirgatus, C. puhuensis, C. roesleri, C. taynguyenensis,
C. teyniei, C. vilaphongi, C. wayakonei from literature
(Ngo and Grismer 2010; David et al. 2004; Hoang et
al. 2007; Heidrich et al. 2007; Ngo and Chan 2011;
Luu et al. 2011; Ulber 1993; Luu et al. 2014; Pauwels et al. 2014; Ngo and Pauwels 2010, Ngo 2011;
Schneider et al. 2011; Ziegler et al. 2003; Rösler et al.
2008; Nguyen et al. 2014; Ziegler et al. 2010; Nguyen
et al. 2013; David et al. 2011; Schneider et al. 2014b;
Nguyen et al. 2010). Due to the high undiagnosed
diversity within the genus, where available, we relied
on examination of topotypic material and/or original
species descriptions.
Molecular comparisons. List of the samples and
sequences with GenBank Accession Numbers is
given in the Table 1. In the molecular analyses we
included 64 samples Cyrtodactylus species from Laos
and central Vietnam. For the Laotian populations
we tried to include several specimens per population
where it was possible. We also included all sequences
on Cyrtodactylus species from Laos, central and
northern Vietnam and southern China available in
the GenBank (published in recent papers: Nguyen
et al. 2013, 2014; Schneider et al. 2014b). COI gene
fragment sequences of a Gekko gecko from southern
China (Wang et al. 2012) was used as outgroup
within the phylogenetic analyses.
DNA extraction, PCR and sequencing. Total
genomic DNA was extracted from ethanol-preserved
muscle or liver tissues using a glass-fiber automatic
DNA isolation protocol following Ivanova et al.
(2006) or using standard phenol–chloroform extraction procedures (Hillis et al. 1996) followed
with isopropanol precipitation. We amplified 673
bp of Cytochrome oxidase I (COI), a mitochondrial
marker proved to be useful for species identification
in reptiles and widely used as a barcoding marker
for vertebrates (Hebert et al. 2003, 2004; Smith et
al. 2008; Solovyeva et al. 2011, 2012; Murphy et al.
2013; Hartmann et al. 2013; Nazarov and Poyarkov
2013), and successfully applied to the studies of
cryptic diversity in the genus Cyrtodactylus (Nazarov et al. 2012; Nguyen et al. 2013, 2014; Schneider
A review of the genus Cyrtodactylus
395
Table 1. Specimens and localities of Cyrtodactylus used in our molecular analysis.
Specimen ID
GenBank
A.N.
Species
Country
Province
Locality
Reference
PNKB4
KF169972
C. cryptus
Vietnam Quang Binh
U Bo, Phong Nha–
Ke Bang NP
Nguyen et al. 2013
PNKB3
KF169971
C. cryptus
Vietnam Quang Binh
U Bo, Phong Nha–
Ke Bang NP
Nguyen et al. 2013
PNKB2
KF169970
C. cryptus
Vietnam Quang Binh
U Bo, Phong Nha–
Ke Bang NP
Nguyen et al. 2013
PNKB1
KF169969
C. cryptus
Vietnam Quang Binh
U Bo, Phong Nha–
Ke Bang NP
Nguyen et al. 2013
ZMMU RAN 1999
HQ543944 C. darevskii sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZIN FN 185
HQ967222 C. darevskii sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZIN FN 188
HQ967225 C. darevskii sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZIN FN 186
HQ967224 C. darevskii sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZIN FN 187
HQ967223 C. darevskii sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZIN FN 256
HQ967221 C. darevskii sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZINFN189
KF929541
C. darevskii sp. nov.
Laos
Khammouane
Na Home, Boulapha
Nguyen et al. 2014
ZINFN187
KF929540
C. darevskii sp. nov.
Laos
Khammouane
Na Home, Boulapha
Nguyen et al. 2014
ZINFN186
KF929539
C. darevskii sp. nov.
Laos
Khammouane
Na Home, Boulapha
Nguyen et al. 2014
ZINFN185
KF929538
C. darevskii sp. nov.
Laos
Khammouane
Na Home, Boulapha
Nguyen et al. 2014
IEBR A.2013.89
KJ817429
C. cf. darevskii sp. nov.
Laos
Khammouane
ITBCZ-10023
KP199951
C. irregularis
ITBCZ-10016
KP199952
ITBCZ-10025
–
Schneider et al. 2014
Vietnam Lam Dong
–
this paper
C. irregularis
Vietnam Lam Dong
–
this paper
KP199953
C. irregularis
Vietnam Lam Dong
–
this paper
ZMMU R-13980-2
KP199946
C. spelaeus sp. nov.
Laos
Vientiane
Khuang Lang N.P., Kasi
this paper
ZMMU R-13980-3
KP199947
C. spelaeus sp. nov.
Laos
Vientiane
Khuang Lang N.P., Kasi
this paper
ZMMU R-13980-1
KP199948
C. spelaeus sp. nov.
Laos
Vientiane
Khuang Lang N.P., Kasi
this paper
ZIN FN 192
HM888468 C. khammouanensis sp. nov. Laos
Khammouane
Na Home, Boulapha
this paper
ZIN FN 257
HM888469 C. khammouanensis sp. nov. Laos
Khammouane
Na Home, Boulapha
this paper
ZIN FN 191
HM888467 C. khammouanensis sp. nov. Laos
Khammouane
Na Home, Boulapha
this paper
ZIN FN257
KF929544
C. khammouanensis sp. nov. Laos
Khammouane
Na Home, Boulapha
Nguyen et al. 2014
ZIN FN192
KF929543
C. khammouanensis sp. nov. Laos
Khammouane
Na Home, Boulapha
Nguyen et al. 2014
ZIN FN191
KF929542
C. khammouanensis sp. nov. Laos
Khammouane
Na Home, Boulapha
Nguyen et al. 2014
IEBR KM2012.52
KP199942
C. lomyenensis
Laos
Khammouane
Lomyen, Gnommalath
this paper
IEBR KM2012.53
KP199943
C. lomyenensis
Laos
Khammouane
Lomyen, Gnommalath
this paper
IEBR KM2012.54
KJ817436
C. lomyenensis
Laos
Khammouane
KIZ201103
KF929537
C. cf. martini
China
Yunnan
–
Xishuangbanna
Schneider et al. 2014
Nguyen et al. 2014
396
R.A. Nazarov et al.
Table 1.( Continued).
Specimen ID
GenBank
A.N.
Species
Country
Province
Locality
Reference
ZMMU RAN 1994-2 HQ967191 C. multiporus sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZMMU RAN 1996-2 HQ967193 C. multiporus sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZIN FN 3
HM888472 C. multiporus sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZIN FN 2
HM888471 C. multiporus sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZIN FN 1
HM888470 C. multiporus sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZMMU RAN 1998
HQ543943 C. multiporus sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZMMU RAN 1995-2
HQ967192 C. multiporus sp. nov.
Laos
Khammouane
Na Home, Boulapha
this paper
ZFMK91827
KJ817431
C. pageli
Laos
Vientiane
PNKN2011119
KF929528
C. phongnhakebangensis
Vietnam Quang Binh
Phong Nha–Ke Bang NP Nguyen et al. 2014
PNKN201132
KF929527
C. phongnhakebangensis
Vietnam Quang Binh
Phong Nha–Ke Bang NP Nguyen et al. 2014
PNKN201130
KF929526
C. phongnhakebangensis
Vietnam Quang Binh
Phong Nha–Ke Bang NP Nguyen et al. 2014
ITBCZ3002
KF169964
C. pseudoquadrivirgatus
Vietnam Thua Thien–Hue
A Luoi
Nguyen et al. 2013
ITBCZ3001
KF169963
C. pseudoquadrivirgatus
Vietnam Thua Thien–Hue
A Luoi
Nguyen et al. 2013
ZMMU R-13095-2
KP199949
C. cf. pseudoquadrivirgatus Vietnam
KIZ11665
KF929529
C. puhuensis
Vietnam Thanh Hoa
IEBR A.2013.109
KJ817428
C. cf. puhuensis
Laos
PNKB201134
KF929532
C. roesleri
Vietnam Quang Binh
Phong Nha–Ke Bang NP Nguyen et al. 2014
PNKB20113
KF929531
C. roesleri
Vietnam Quang Binh
Phong Nha–Ke Bang NP Nguyen et al. 2014
PNKB20111
KF929530
C. roesleri
Vietnam Quang Binh
Phong Nha–Ke Bang NP Nguyen et al. 2014
IEBR A.2013.112
KJ817437
C. cf. roesleri
Laos
ROM32120
KF169979
C. taynguyenensis
Vietnam Gia Lai
Krong Pa, K Bang
Nguyen et al. 2013
ROM32119
KF169978
C. taynguyenensis
Vietnam Gia Lai
Krong Pa, K Bang
Nguyen et al. 2013
IEBR KM2012.14
KP199944
C. teyniei
Laos
Khammouane
–
this paper
IEBR KM2012.77
KP199945
C. teyniei
Laos
Khammouane
–
this paper
IEBR KM2012.77
KJ817430
C. teyniei
Laos
Khammouane
–
Schneider et al. 2014
NUOL R-2013.5
KJ817434
C. vilaphongi
Laos
Luang Prabang
–
Schneider et al. 2014
IEBR A.2013.103
KJ817435
C. vilaphongi
Laos
Luang Prabang
–
Schneider et al. 2014
ZMMU R-13981-1
KP199950
C. wayakonei
Laos
Luang Nam Tha
–
this paper
ZFMK91016
KJ817438
C. wayakonei
Laos
Luang Nam Tha
–
Schneider et al. 2014
ITBCZ2532
KF169962
Cyrtodactylus sp. 1
Vietnam Da Nang
IEBR A.2013.110
KJ817432
Cyrtodactylus sp. 2
Laos
Luang Prabang
–
Schneider et al. 2014
IEBR A.2013.111
KJ817433
Cyrtodactylus sp. 2
Laos
Luang Prabang
–
Schneider et al. 2014
JF920657
Gekko gecko
China
–
Wang et al. 2012
–
–
–
–
Pu Hu
Houphan
–
this paper
Nguyen et al. 2014
–
Khammouane
Schneider et al. 2014
–
Ba Na
Schneider et al. 2014
Schneider et al. 2014
Nguyen et al. 2013
A review of the genus Cyrtodactylus
397
Fig. 1. General distribution of the genus Cyrtodactylus in Central Annamites and surrounding areas; dot in the center of icon indicates the
type locality; large dot indicates type localities for several species of Cyrtodactylus: 1 – Cyrtodactylus cf. martini – Xishuangbanna, Yunnan
Province, China; 2 – C. martini – Lai Chau, Lai Chau Province, Vietnam; 3 – C. bichnganae – Son La, Son La Province, Vietnam; 4 – C.
wayakonei – Kao Rao Cave, Ban Nam Eng, Vieng Phoukha, Luang Nam Tha Province, Laos; 5 – C. huongsonensis – Huong Son, My Duc,
Hanoi, Vietnam; 6 – C. cucphuongensis – Cuc Phuong N.P., Nho Quan, Ninh Binh Province, Vietnam; 7 – С. puhuensis – Pu Hu, Thanh Hoa
Province, Vietnam; 8 – C. cf. puhuensis – Houphan Province, Laos; 9 – C. vilaphongi – Ban Xieng Muak, Luang Prabang, Luang Prabang
Province, Laos; 10 – C. chauquangensis – Chau Quang, Quy Hop, Nghe An Province, Vietnam; 11 – C. cf. interdigitalis and C. spelaeus sp.
nov. – Khuang Lang N.P., Kasi, Vientiane Province, Laos; 12 – C. pageli – Phoukham Cave, Ban Na Thong, Vang Vieng, Vientiane Province, Laos; 13 – C. teyniei – Ban Na Hin, Nam Kading NBCA, Borikhamxay Province, Laos; 14 – C. jarujini – Phu Wua W.S., Nong Dern,
Bung Kan, Nong Khai Province, Thailand; 15 – C. cf. teyniei –Nahin, Khammouane Province, Laos; 16 – C. lomyensis – Lomyen Cave,
Gnommalath, Khammouane Province, Laos; 17 – C. darevskii sp. nov., C. khammouanensis sp. nov. and C. multiporus sp. nov.– Na Home,
Boulapha, Khammouane Province, Laos; 18 – C. phonghnakebangensis, C. cryptus and C. roesleri – Phong Nha – Ke Bang N.P., Minh Hoa,
Quang Binh Province, Vietnam; 19 – C. jaegeri – Thakhek, Khammouane Province, Laos; 20 – C. kunyai – Suan Hin Pha Ngam, Nong
Hin District, Loei Province, Thailand; 21 – C. interdigitalis – Tham Yai Nam Nao, Nam Nao N.P., Petchabun Province, Thailand; 22 – C.
pseudoquadrivirgatus – Huong Hoa, Quang Tri Province, Vietnam; 23 – C. pseudoquadrivirgatus – A Luoi, Thua Thien – Hue Province,
Vietnam; 24 – Cyrtodactylus sp. 1 – Ba Na, Da Nang, Vietnam; 25 – C. buchardi – Kiatngong, Xepian NBCA, Champasak Province, Laos;
26 – C. cf. taynguyenensis – Kon Plong, Kon Tum Province, Vietnam; 27 – C. taynguyenensis – Krong Pa, Gia Lai Province, Vietnam.
398
et al. 2014a, 2014b). Primers used both for PCR
and sequencing were the VF1-d (5’-TTCTCAACCAACCACAARGAYATYGG-3’) and the VR1-d
(5’-TAGACTTCTGGGTGGCCRAARAAYCA-3’)
(following Ivanova et al. 2006). The obtained fragments were sequenced in both directions for each
sample, and a consensus sequence was generated.
PCRs were performed in 25 μl reactions using ca.
50 ng genomic DNA, 10 pmol of each primer, 15
nmol of each dNTP, 50 nmol additional MgCl 2, Taq
PCR buffer (10 mM Tris-HCl, pH 8.3, 50 mM KCl,
1.1 mM MgCl 2 and 0.01% gelatine) and 1 U of Taq
DNA polymerase. The PCR conditions were: an initial denaturation step at 95 °C for 3 min; 5 cycles at
95 °C for 30 s, annealing at 45 °C for 1 m, extension
at 72 °C for 2 min followed with 35 cycles at 95 °C
for 30 s, annealing at 51 for 1 m, extension at 72 °C
for 2 min and final extension of 5 min at 72 °C. PCR
products were loaded onto 1% agarose gels, stained
with GelStar gel stain (Cambrex), and visualized in
a Dark reader transilluminator (Clare Chemical). If
results were satisfying, products were purified using
2 μl, from a 1:4 dilution of ExoSapIt (Amersham), per
5 μl of PCR product prior to cycle sequencing. A 10 μl
sequencing reaction included 2 μL of template, 2.5 μl
of sequencing buffer, 0.8 μl of 10 pmol primer, 0.4 μl
of BigDye Terminator version 3.1 Sequencing Standard (Applied Biosystems) and 4.2 μl of water. The
sequence reaction was 35 cycles of 10 sec at 96 °C, 10
s at 50 °C and 4 min at 60 °C. Cycle sequencing products were purified by ethanol precipitation. Sequence
data collection and visualization were performed on
an ABI 3730xl automated sequencer (Applied Biosystems). Obtained sequences are accessible at BOLD
systems website (http://www.boldsystems.org) and
are deposited in GenBank under accession numbers
HM888467–HM888472; HQ543943–HQ543944;
HQ967191–HQ967193; HQ967221–HQ967225;
KP199942–KP199953 (see Table 1).
Phylogenetic analysis. Final alignment used
for phylogenetic analysis contained 673 bp of COI
gene for 64 specimens belonging to approximately
19 Cyrtodactylus species and 1 outgroup sequence
of Gekko gecko (summarized in Tabl. 1). Sequences
were initially aligned automatically by ClustalX
1.81 (Thompson et al. 1997) and then optimized
by eye using BioEdit Sequence Alignment Editor
5.0.9 (Hall 1999). Sequences were also submitted
to a BLAST search in GenBank to confirm that the
intended sequences had been amplified. Mean un-
R.A. Nazarov et al.
corrected genetic distances (p-distances) between
sequences were determined with MEGA 6.0 (Tamura
et al. 2013). MODELTEST v.3.06 (Posada and Crandall 1998) was used to estimate the optimal evolutionary models to be used for the data set analysis.
The best-fitting model as suggested by the Akaike
Information Criterion (AIC) was the (GTR + I + G)
model of DNA evolution. Phylogenetic analyses were
conducted in PAUP version 4.0b4a (Swofford 1998)
and MEGA6.06 (Tamura et al. 2013) software.
Phylogenetic analyses were conducted in
Treefinder (Jobb et al. 2004) and MrBayes 3.1.2
(Huelsenbeck, Ronquist 2001; Ronquist, Huelsenbeck 2003) software. The Maximum Likelihood
(ML) analyses were conducted using Treefinder
(Jobb et al. 2004). Transitions and transversions were
equally weighted, and gaps were treated as missing
data. Confidence in tree topology was tested by nonparametric bootstrap analysis (Felsenstein 1985)
with 1000 replicates, and by posterior probability
(PP) for Bayesian inference (BI) in MrBayes 3.1.2
(Huelsenbeck, Ronquist 2001). We a priori regarded
tree nodes with bootstrap values 70% or greater and
posterior probabilities values over 0.95 as sufficiently
resolved, those between 75% and 50% (0.95 and 0.90
for BI) were regarded as tendencies, those below 50%
(0.90 for BI) were considered to be non-resolved
(Huelsenbeck and Hillis 1993).
RESULTS
Molecular differentiation of Laotian Cyrtodactylus
Sequence statistics. The final alignment of the
studied COI mtDNA fragment consisted of 673 sites
in which 407 sites were conserved, 266 variable and
258 of them were potentially parsimony-informative;
the transition–transversion bias was estimated as
2.49 (all data given for ingroups only). Substitution
rates were estimated under the Kimura’s 2-parameter
model (+G+I). Nucleotide frequencies were A =
23.0%, T = 26.0%, C = 31.0%, and G = 20.0%.
Sequence divergence. The uncorrected p-distances among and within COI gene fragment sequences
of the studied Cyrtodactylus taxa and the outgroup
(Gekko gecko) are shown in the Table 2.
Phylogenetic relationships. As shown by a
number of previous studies, phylogenetic analysis
of the COI fragment is a powerful tool for molecular
diagnostics of biodiversity within the taxonomi-
A review of the genus Cyrtodactylus
cally complicated genus Cyrtodactylus (Nazarov et al.
2012; Nguyen et al. 2013, 2014; Schneider et al. 2011,
2014a, 2014b). Though resolving phylogeny of the
genus Cyrtodactylus would require more genes and
taxa to be studied (see Wood et al. 2012), the 673-bp
fragment of the COI analyzed herein demonstrates
phylogenetic signal which is strong enough to provide significant support values for some nodes in the
resulted tree. These phylogenetic patterns are briefly
discussed below.
The results of the phylogenetic analysis of the
COI gene are shown in Fig. 2. Phylogenetic relationships between the accessed taxa of Cyrtodactylus are
poorly resolved with major basal nodes in the tree
having low (BS < 75%) or insignificant levels (BS <
50%; BPP < 0.95) of support, whereas monophyly of
species-level groups and species complexes is significantly supported (BS > 90%; BPP 0.95).
Both ML and BI analyses resulted in essentially
similar topologies. They differed only in associations at poorly supported nodes. The ML tree (Fig.
2) infers the following set of phylogenetic relationships among studied Cyrtodactylus species, which in
general corresponds well to the preliminary tree for
Laotian species reported by Schneider et al. (2014b):
(1) The phylogenetic analyses revealed 21 lineages mtDNA haplotypes in Cyrtodactylus of Laos
and adjacent parts of Vietnam and China included in
the present work (see groups 1–21 in Fig. 2). These
groups roughly correspond to species-level of differentiation and have high levels of BS and PP statistical support of their monophyly.
(2) Altogether, the revealed 21 matrilineal lineages of Cyrtodactylus are grouped into three major
clades, corresponding to the level of species groups
in the genus Cyrtodactylus (see groups A–C in Fig.
2). The level of statistical support of their monophyly
varies from high (for groups A and C) to moderate
(for group B).
(3) Group A, which we here tentatively indicate
as C. phongnhakebangensis species group, includes
lineages 1–9 which represent species associated with
limestone habitats in central Vietnam (C. phongnhakebangensis (2, indicates lineage number in Fig. 2
and Tabl. 2), C. roesleri (3)) and central and eastern
Laos (C. lomyenensis (6) and C. teyniei (7) from
Khammouane Province, C. pageli (9) from Vientiane
Province). The latter species, C. pageli (lineage 9) appears to be more distantly related to others, though
its’ phylogenetic position is recovered in alternative
399
ways in ML and BI analyses (always with non-significant values of node support).
(4) The sample IEBR A.2013.112 from Khammouane Province in Laos, indicated as “Cyrtodactylus
sp. 4” in the original publication by Schenider et al.
(2014b) with absolute values of node support (BS/
PP = 100/1.0) groups with samples of C. roesleri from
Quang Binh Province, Vietnam (lineage 4 – C. cf.
roesleri in Fig. 2). Certain level of genetic differentiation between this lineage and C. roesleri from the type
locality (p = 6.12%, see Tabl. 2) may be explained by
incomplete taxonomy of this group as well as by interspecific differentiation of local populations. Thus, our
analysis confirms the presence of C. roesleri or a very
closely related form of Cyrtodactylus in Laos; herein
we tentatively indicate this lineage as C. cf. roesleri,
further morphological examination and analysis of
nuclear DNA markers might help to clarify the taxonomic status of this lineage.
(5) The three syntopic morpho-species of Cyrtodactylus found by us in limestone forests in the
environs of Na Home, Khammouane Province, eastern Laos, group into three distantly related clades;
all belonging to the C. phongnhakebangensis species
group. Among them, clade 1 includes the sample
IEBR A.2013.89, indicated as “Cyrtodactylus sp. 3” in
the original publication by Schenider et al. (2014b),
genetic distance from our samples is less than p =
1.0% (see Tabl. 2). This species is recovered as a sistergroup of C. phongnhakebangensis from Quang Binh
Province of Vietnam (see Fig. 2); it was indicated as
“Cyrtodactylus sp. 3” (Schenider et al. 2014b) or as
“Cyrtodactylus sp. 4” (Nguyen et al. 2014; please note
that sample IDs and GenBank AN’s are erroneously
mixed in the Table 1 of this publication) by previous
researchers. The uncorrected genetic distinace of the
clade 1 from its sister species C. phongnhakebangensis
reaches p = 9.34% (see Tabl. 2). Based on significant
genetic distance in COI gene and morphological differences, below we describe the representatives of this
clade as a new species, Cyrtodactylus darevskii sp. nov.
(6) The clade 5 also from Na Home Village in
Khammouane Province of Laos forms a clearly distinct mtDNA lineage (see Fig. 2). This species was
also included in the recent paper of Nguyen et al.
(2014) where it was indicated as “Cyrtodactylus sp. 5”
(Nguyen et al. 2014; please note that sample IDs and
GenBank AN’s are erroneously mixed in the Table 1
of this publication). Genetic differentiation of this
lineage from other members of the C. phongnhakeban-
Species
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
1. C. darevskii sp. nov.
0.99
1.14
1.66
1.54
1.45
1.52
1.46
1.53
1.40
1.63
1.72
1.63
1.79
1.62
1.61
1.66
1.59
1.65
1.68
2.19
1.73
1.72
2. C. phongnhakebangensis
9.34
0.00
1.60
1.64
1.50
1.54
1.42
1.52
1.54
1.66
1.76
1.75
1.71
1.63
1.67
1.68
1.67
1.90
1.71
2.13
1.77
1.64
3. C. roesleri
17.23 15.33
0.24
1.11
1.68
1.69
1.46
1.53
1.65
1.53
1.60
1.70
1.69
1.78
1.78
1.74
1.65
1.50
1.78
2.35
1.81
1.73
4. C. cf. roesleri
15.93 16.36
6.12
—
1.67
1.71
1.49
1.52
1.61
1.51
1.56
1.75
1.57
1.79
1.75
1.70
1.65
1.46
1.77
2.36
1.91
1.82
5. C. khammouanensis sp. nov.
14.93 15.54 16.42 16.69
0.10
1.49
1.51
1.55
1.46
1.47
1.50
1.67
1.61
1.61
1.57
1.65
1.66
1.75
1.63
2.10
1.57
1.75
6. C. lomyenensis
14.56 14.69 17.67 17.97 15.60
0.12
1.30
1.35
1.33
1.47
1.55
1.79
1.62
1.65
1.56
1.63
1.60
1.65
1.71
2.04
1.65
1.75
7. C. teyniei
15.34 15.27 17.52 17.82 17.18 14.57
0.00
0.86
1.50
1.56
1.46
1.64
1.62
1.64
1.55
1.71
1.64
1.56
1.76
2.32
1.64
1.70
8. C. multiporus sp. nov.
15.45 15.27 16.97 17.09 16.57 14.39
6.00
0.00
1.56
1.54
1.45
1.65
1.62
1.65
1.57
1.69
1.58
1.59
1.77
2.35
1.68
1.65
9. C. pageli
17.74 17.64 17.52 16.73 18.59 17.06 17.64 17.45
—
1.41
1.41
1.69
1.45
1.60
1.62
1.72
1.70
1.57
1.91
2.01
1.57
1.70
10. C. puhuensis
22.88 21.45 19.73 20.09 22.09 21.10 20.91 21.36 20.09
3.27
0.96
1.33
1.49
1.57
1.52
1.38
1.33
1.44
1.49
1.77
1.45
1.77
11. C. vilaphongi
20.48 20.55 20.55 20.00 21.85 20.95 18.36 19.64 19.27
9.45
0.00
1.27
1.47
1.51
1.51
1.39
1.50
1.43
1.55
1.91
1.52
1.67
12. C. spelaeus sp. nov.
23.53 20.79 20.00 20.00 21.65 23.38 21.03 21.21 21.45 11.48 12.30
0.61
1.49
1.51
1.50
1.61
1.49
1.68
1.71
1.97
1.68
1.82
13. C. sp. 2 Luang Prabang
20.71 18.73 20.24 19.27 19.32 19.98 21.64 21.27 20.36 15.00 13.45 14.48
0.00
1.49
1.57
1.49
1.62
1.56
1.50
1.91
1.65
1.70
14. C. martini
21.82 18.91 20.48 20.91 20.26 22.04 22.36 22.18 19.45 17.18 15.45 15.39 14.55
—
0.97
1.58
1.57
1.70
1.60
2.03
1.53
1.62
15. C. wayakonei
21.05 19.91 21.58 20.91 20.54 21.40 22.45 22.27 20.55 18.23 16.55 16.67 16.09
6.82
0.91
1.64
1.50
1.71
1.61
2.07
1.67
1.68
16. C. cryptus
20.61 19.64 22.24 20.73 20.69 20.58 20.36 20.36 19.64 20.00 18.73 19.94 20.55 19.27 18.55
0.00
1.23
1.40
1.46
1.78
1.41
1.88
17. C. sp. 1 Da Nang
22.36 20.36 21.15 21.82 22.55 21.31 20.18 19.64 20.00 18.73 17.82 19.39 19.09 17.45 18.18
9.27
—
1.44
1.31
1.79
1.41
1.68
18. C. irregularis
22.24 21.39 22.12 20.97 22.91 21.62 21.39 22.30 21.52 20.52 18.42 22.06 19.52 19.88 19.52 16.06 15.52
0.12
1.54
1.81
1.36
1.79
19. C. taynguyenensis
21.64 21.09 23.27 21.45 20.87 20.58 20.36 22.00 22.36 20.00 19.27 20.55 21.09 19.64 18.82 15.09 15.82 16.61
0.00
1.42
1.24
1.79
20. C. cf. pseudoquadrivirgatus
22.75 19.73 22.07 22.70 21.08 20.33 21.89 21.62 20.54 19.59 19.73 20.81 19.46 17.57 18.92 15.68 13.24 13.78 10.27
—
1.37
2.19
21. C. pseudoquadrivirgatus
20.63 19.82 21.15 22.00 20.20 21.74 20.00 21.64 20.91 18.36 18.36 19.58 21.27 17.27 17.82 13.27 14.55 13.70 12.73
8.11
0.00
1.67
22. Gekko gecko
23.09 22.36 23.09 23.27 22.67 22.89 24.91 23.82 26.00 23.09 24.00 24.61 23.09 22.91 23.36 24.00 24.18 25.39 25.64 24.86 25.09
—
R.A. Nazarov et al.
1
400
Table 2. Uncorrected p-distance (percentage) between COI sequences of studied Cyrtodactylus species (below diagonal), within group mean p-distances (on diagonal) and
calculation errors (above diagonal).
A review of the genus Cyrtodactylus
401
Fig. 2. ML-tree of studied Cyrtodactylus species based on the analysis of 673 bp of COI mtDNA gene. Node colour indicates support value:
black for well-supported and sufficiently supported nodes (ML BS > 90%; BI PP > 0.95), greyfor moderately or poorly supported nodes
and white for unresolved nodes with low or no support. Bootstrap support (BS) and posterior probability (PP) values are shown above or
below tree nodes for ML/BI analyses respectively. Genbank accession numbers of samples corresponds to those in Table 1.
402
gensis species group is significant, with minimal p-distances found between this species and C. roesleri from
Quang Binh Province of Vietnam and Khammouane
Province of Laos (p = 16.42–16.69%, see Tabl. 2).
Based on high genetic distances between COI gene
sequences and significant morphological differences,
below we describe the members of this clade as a new
species, Cyrtodactylus khammouanensis sp. nov.
(7) The members of the clade 8 from environs of
Na Home Village in Khammouane Province of Laos
form a significantly supported monophyletic group
with another Laotian species C. teyniei (see Fig. 2),
which also inhabits limestones of Khammouane Province (see Fig. 1). These lineages appear to be very close
with minimal uncorrected genetic distance observed
between them: p = 6.00%. The genetic distances of
this lineages from another limestone species from the
Khammouane Province, C. lomyenensis, is much grater (p = 14.39–14.57%, see Tabl. 2). However, stable
differences in unique states of morphological features
(see below) allow us to designate this lineage as a new
species, Cyrtodactylus multiporus sp. nov.
(8) Monophyly of the group B, encompassing
limestone and cave species of Cyrtodactylus from central, western and northern Laos and adjacent China,
is moderately supported (BS / PP = 74/0.94); tentatively we indicate it as C. wayakonei species group
(see Fig. 2). This group is subdivided into two subclades, one joining C. wayakonei from northern Laos
and Cyrtodactylus sp. from Xishuangbanna (which,
based on preliminary examination of vouchers, we
tentatively indicate here as C. cf. martini) and the second one, joining all other species. Among the latter,
the sample Cyrtodactylus from Houphan Province in
Laos (IEBR A.2013.109) indicated as “Cyrtodactylus
sp. 1” in the original publication by Schenider et al.
(2014b) is undoubtedly with absolute levels of statistical support (BS/PP = 100/1.0) conspecific with
the recently described C. puhuensis from Thanh Hoa
Province of Vietnam (see Fig. 2). Genetic differentiation between Laotian and Vietnamese populations
of this species is minimal (p = 3.27%; see Tabl. 2);
thus our analysis provides evidence for including C.
puhuensis in the herpetofaunal list of Laos. The species appears to be closely related to C. vilaphongi from
Luang Prabang Province in north-central Laos (p =
9.45%, see Tabl. 2).
(9) The species indicated as “Cyrtodactylus sp. 2”
in the original publication by Schenider et al. (2014b)
appears to be quite distant from other members of the
R.A. Nazarov et al.
group (p = 13.45–15.00%, see Tabl. 2). The population of cave-dwelling Cyrtodactylus from Khuang
Lang N.P., Kasi District, Vientiane Province, also
appears to be quite distant from other members of
C. wayakonei species group (p = 11.48–11.48%).
Based on distinct morphological differences and significant differentiation in mtDNA we below describe
this species as Cyrtodactylus spelaeus sp. nov.
(10) The members of the clade C belong to the
C. irregularis species group, which in our analysis
includes only central-Vietnamese taxa: C. cryptus
from Quang Binh Province, Cyrtodactylus sp. 1 from
Da Nang, C. irregularis from Lam Dong Province,
C. taynguyenensis from Gia Lai Province, C. cf. pseudoquadrivirgatus and C. pseudoquadrivirgatus from
Thua Thien – Hue Province. Phylogenetic relationships between these closely-related taxa in general
correspond to those reported by previous researchers
(Nazarov et al. 2012; Nguyen et al. 2013, 2014).
Taxonomic accounts
Cyrtodactylus darevskii sp. nov.
(Fig. 3)
Holotype. Adult male ZMMU R-13980 (field
number F185) collected on 18 June 2009 by Nikolai Orlov, Sang Ngoc Nguyen and Konstantin Milto
in environs of Na Phao Village, Boulapha District,
Khammouane Province, Laos (17°34´57.1´´N and
105°44´37.3´´E; elevation 170 m a.s.l.).
Paratypes. Three adult males (ZMMU R-139811; 13981-2 (FN 186, 187); ZIN 28247 (FN 256), two
adult females (ZMMU 13981-3 (1999); ZIN 28248
(FN 188) and one subadult ZIN 28249 (FN 189). All
members of the type series with the same collection
data as the holotype.
Diagnosis. A large sized, slender Cyrtodactylus
with a maximum SVL of 100 mm; the new species
is distinguished from all congeners by the following morphological characters. Dorsal color pattern
consisting of narrow dark nuchal band and 4–5 dark
transversal breaking bands with light yellowish edge
between limbs and 8–10 transversal dark bands on
the tail. Dorsal surface of head very light brown,
sometimes with a yellowish tinge, with few small
irregular roundish dark brown spots. Dorsal of head
and temporal region, body, hind limbs and base of tail
covered by rounded, keeled tubercles, which are 2–3
times larger than the surrounding scales. Ventrals in
A review of the genus Cyrtodactylus
403
Fig. 3. Cyrtodactylus darevskii sp. nov.: A – general view of holotype; B – precloacal region with continuous row of femoral and precloacal
pores; C – dorsum surface with the rows of enlarged tubercles; D – dorsal surface of the head; E – ventral surface of the head with mental
scalation.
404
38–46 longitudinal rows at midbody, lateral folds are
not strongly developed. Continuous series of 38–44
precloacal and femoral pores in males and 24–34
precloacal and femoral pores in females. Subcaudals
in transversally enlarged median row, flat, smooth,
imbricate. No enlarged keeled tubercles on the dorsal
surface of the tail; 4–5 postcloacal spurs in both sexes.
Holotype description. The holotype is an adult
male, in good state of preservation; it has the following measurements: SVL 90.0 mm, TailL 105.0 mm,
HeadL 24.8 mm, HeadW 15.8 mm, HeadH 9.7 mm ,
SnEye 10.5 mm, OrbD 5.5 mm, EarL 2.2 mm, EyeEar
6.2 mm; proportions are as follows: SVL/HeadL 3.62,
HeadL/HeadW 1.56, HeadL/HeadH 2.55, SnEye/
EyeEar 1.69.
Rostral is large, wider than high (RW 3.9 mm, RH
2.2 mm, RW/RH 1.77) with an inverse Y–shaped
median suture; supralabials 10/10; infralabials 9/10;
nares surrounded by rostral anteriorly, first supralabial laterally, supranasal and two nasals posteriorly;
rostral about 4–5 times larger than supranasal; supranasals in broad contact with each other; upper anterior ciliaries two times larger than posterior ones;
head scales granular, two times smaller than the median snout scales; no enlarged keeled tubercles on the
dorsal surface of head (Fig. 3d); on temporal region
enlarged tubercles are present, they are 3–4 times
larger than the surrounding scales; mental triangular,
as wide as rostral; two pairs of enlarged postmentals,
first pair longer than wide and in broad contact (Fig.
3e); dorsal scales granular, 3–4 times smaller than
the ventral scales; dorsal tubercles round, flat, keeled,
surrounded by 8–10 granular scales, tubercles forming about 18 irregular longitudinal rows at midbody
(Fig. 3c); ventral scales smooth, 38 longitudinal rows
at midbody; lateral folds weakly developed; dorsal
surface of fore limbs without enlarged tubercles and
hind limbs with smooth roundish enlarged tubercles;
fingers and toes without web, basal lamellae more
rounded than on distal surface of digits; row of enlarged femoral and precloacal scales with 43 pores,
(Fig. 3b); fore pairs of enlarged postcloacal spurs;
dorsal of tail without enlarged keeled tubercles; posterior part of tail covered by flattened and rounded
scales; subcaudals with enlarged median plate row,
flat, smooth, imbricate, twice wider than high.
Coloration: Dorsal surface of head is light brown
with the few roundish small dark irregular dots; nuchal band not wide with undulating edge posteriorly,
dark brown with a distinct light margin, extending
R.A. Nazarov et al.
from the neck to the posterior margins of eyes; labials are light grey. Dorsal color pattern formed by five
transversal dark brown bands with light yellowish
edge between limbs. Dorsal surface of limbs and digits light brown or yellowish with unclear dark bands.
The lower surfaces of toes and fingers are light grey.
Tail with eight, wide, dark brown bands, which are
wider than the light grey interceptions in-between.
Ventral body surfaces white, tail dark with light
spots. For coloration in life see Fig. 3a.
Variation of paratypes. For the variation of the
type series see Table 3. The dorsal pattern is somewhat variable. Sexual dimorphism is not well developed, males somewhat smaller than females, maximal
SVL for males is 93.2 mm and for females is 100 mm.
Precloacal and femoral pores present in both sexes,
but males have more developed postcloacal spurs.
Comparisons with Laotian congeners. Below
we compare the new species with eleven Laotian
congeners. Cyrtodactylus darevskii sp. nov. differs
from C. buchardi David, Teynie et Ohler, 2004 by
the singe median row of subcaudals (vs. subcaudals
not enlarged in C. buchardi), enlarged femoral scales
(lacking in C. buchardi), fewer dorsal tubercle rows
(16–20 vs. 25), more ventral scales (38–46 vs. 30),
and more subdigital lamellae under the fourth finger
and toe (17–20 and 18–22 vs. 14 and 12).
From C. interdigitalis Ulber, 1993 new species differs by transversal enlarged subcaudal scales; higher
number of precloacal and femoral pores in the singe
row (38–44 for males and 24–38 for females vs. 14
precloacal and 9+9 femoral pores for C. interdigitalis);
roundish tail versus flattened tail; dorsal patterns of
new species consisting of 4–5 dark transverse narrow
bands versus 4–5 wide brownish jagged transversal
bands; no webbing between toes versus developed
web on the basis of toes. Cyrtodactylus darevskii sp.
nov. can be distinguished from C. jaegeri Luu, Calame, Bonkowski, Nguyen et Ziegler, 2014 by larger
body size (maximum SVL 100 mm vs. 68.5 mm), a
higher number of ventral scales (38–46 vs. 31–32),
dorsal color patterns (dark narrow transverse wavy
bands vs. wide transversal dark bands). The new
species is distinguished from C. jarujini Ulber, 1993
by having smaller number of femoral and precloacal
pores arranged in a continuous row (38–44 vs. 52–54
pores in an irregular row), more ventral scales (38–46
vs. 30–38), and also can be further distinguished by
number of subdigital lamellae (LF4 17–20 and LT4
18–22 vs. 12–17 and 11–18 respectively). The new
A review of the genus Cyrtodactylus
405
Table 3. Measurements and selected morphological characters of the type series of Cyrtodactylus darevskii sp. nov.; f = female, m= male,
sub = subadult.
Holotype
Paratypes
ZMMU
R-13980
ZMMU
R-13981-2
ZMMU
R-13981-1
ZIN 28247
ZMMU
R-13981-3
ZIN 28248
ZIN 28249
m
m
m
m
f
f
sub
SVL
90
93.2
90
73.7
100
84.6
50.6
TailL
105
*
113
95
*
45*
*
15.0
Sex
Head L
24.8
25.3
24.5
21.5
27.6
23.2
Head W
15.8
15.8
16
13
17.7
14.2
9.0
Head H
9.7
9.4
10
7.7
10.8
8.7
5.2
SnEye
10.5
10.6
10.3
8.6
11.2
9.0
4.7
OrbD
5.5
5.7
5.2
4.8
5.8
5.0
3.9
EarL
2.2
2.6
1.8
2.0
2.7
2.5
1.0
EyeEar
6.2
5.7
6.2
5.3
6.5
5.7
3.6
TrunkL
40.1
39.5
38.2
32.6
44.3
38
19.8
LS
15.5
16.1
15.3
12.9
16.7
14.2
7.8
ForeaL
14
14.8
14.4
11.5
15.3
13.2
7.2
FemurL
18.5
21
17.7
16.6
21.2
18.3
10.7
Crus L
14.7
17
15.3
12
16.2
13.8
8.3
LD4A
9.1
9.2
9.0
8.0
9.6
7.7
4.6
LD4P
9.8
12.4
11.7
9.7
12.0
10.5
5.7
V
38
43
40
40
44
46
40
SLB
193
194
216
180
192
208
190
LF 4
19
17
20
19
19
19
17
18
LT 4
20
18
20
18
22
20
PP+FP
43
44
38
38
34
24
0
SL
10
12
10
11
11
10
12
IL
9
10
10
9
10
9
11
TubL
42
43
43
40
44
48
38
TubW
18
16
16
20
16
18
18
SLH
93
97
95
85
90
91
94
SAH
78
72
82
71
74
75
80
species is distinguishable from C. lomyenensis Tri et
Pauwels, 2010 by the larger body size (maximum
SVL 100 mm vs. 72.1 mm in C. lomyenensis), a higher
number of ventral scales (38–46 vs. 35–36), dorsal
color pattern (narrow dark transverse bands vs. wide
bands in C. lomyenensis). Cyrtodactylus darevskii
sp. nov. can be further differentiated from C. pageli
Schneider, Nguyen, Schmitz, Kingsada, Auer et
Ziegler, 2011 by the following morphological attributes: continuous row of precloacal and femoral pores
(38–44 vs. 4–6 precloacal pores in C. pageli), greater
number of enlarged dorsal tubercle rows (16–20 vs.
9–14)*. From C. roesleri Ziegler, Nazarov, Orlov,
Nguyen, Vu, Dang, Dinh et Schmitz, 2010 the new
*The number of pores in the single known female is ambiguously stated in the original description of C. teyniei; in Tab.
1 (p. 37, David et al. 2011) the number of pores is given as 13, whereas in the diagnosis on the page 30 and in the Fig. 3a of
the same publication the number of pores is stated as 14.
406
species is distinguishable by having a higher number
of precloacal and femoral pores (38–44 vs. 20–28),
larger maximum body size (100 mm vs. 75.3 mm),
dorsal pattern (irregular, dark transverse bands vs.
wide transverse dark bands in C. roesleri). The new
species differs from C. teyniei David, Nguyen, Schneider et Ziegler, 2011, by the higher number of pores
in females (24–34 vs. 13 or 14 ), dorsal color pattern
(narrow bands vs. blotches in C. teyniei), presence of
nuchal loop (present in the new species vs. absent in
C. teyniei). The new species can be further diagnosed
from C. puhuensis Nguyen, Yang, Thi Le, Nguyen,
Orlov, Hoang, Nguyen, Jin, Rao, Hoang, Che, Murphy et Zhang, 2014 by a higher number of precloacal
and femoral pores (38–44 vs. 5 precloacal pores in C.
puhuensis), and by a different dorsal pattern (dark
transverse bands vs. light narrow bands in C. puhuensis). From C. vilaphongi Schneider, Nguyen, Duc Le,
Nophaseud, Bonkowski et Ziegler, 2014 the new
species differs by having enlarged median row of subcaudals (absent in C. vilaphongi), higher number of
ventral scales (38–46 vs. 34–36 in C. vilaphongi) and
different dorsal color pattern (dark transverse bands
vs. narrow yellowish white bands in C. vilaphongi).
From C. wayakonei Nguyen, Kingsada, Roesler, Auer
et Ziegler, 2010 the new species is distinguished by a
higher number of precloacal and femoral pores (38–
44 vs. 6–8 precloacal pores), different dorsal pattern
(dark bands vs. blotched to reticulated pattern in C.
wayakonei), and by subcaudal scalation (enlarged
median row of subcaudals vs. somewhat enlarged and
broadened subcaudals).
Etymology. Cyrtodactylus darevskii sp. nov. is
named for the honor of the famous Russian herpetologist Ilya Sergeevich Darevsky (1924–2009).
The recommended vernacular name in English:
Darevsky’s Bent-toed Gecko.
Habitat description. The study area is located
in environs of Na Phao Village, Boulapha District,
Khammouane Province, eastern Laos. (17°34´57.1´´N
and 105°44´37.3´´E, elevation 170 m a.s.l.). Field
work was conducted there during 15–20 June,
2009. The study area is situated near Phou Hinboun
(Limestone) National Biodiversity Conservation
Protected Area. The area is dominated by sparsely
vegetated limestone karst. Cyrtodactylus darevskii
sp. nov. was found at the altitude 170 m a.s.l. in a
karst cave (Fig. 8). All the geckos are found on the
walls in small-sized caves. They are recorded at night
near numerous cracks that serve as shelters. The new
R.A. Nazarov et al.
species was recorded in synbiotopy with Cyrtodactylus khammouanensis sp. nov. (see below).
Distribution. To date the new species is known
only from the type locality.
Phylogenetic position. A member of C. phongnhakebangensis species group; most closely related to
its sister species C. phongnhakebangensis, distributed
in Quang Binh Province of Vietnam (p = 9.34%).
Cyrtodactylus khammouanensis sp. nov.
(Fig. 4)
Holotype. Adult male ZMMU R-13982 (field
number FN 192) collected on 16 June 2009 by Nikolai Orlov, Sang Ngoc Nguyen and Konstantin Milto
in the environs of Na Phao Village, Boulapha District, Khammouane Province, Laos (17°34´57.1´´N
and 105°44´37.3´´E, elevation 170 m a.s.l.).
Paratypes. One adult male ZIN 28250 (FN 193)
and two adult females ZMMU R-13983 (FN 257);
ZIN 28251 (FN 191). All type specimens are with the
same data as the holotype.
Diagnosis. Relatively small Cyrtodactylus with a
maximum SVL of 73 mm; the new species is distinguished from all other congeners by the combination
of the following morphological characters. Dorsal
pattern consisting of wide dark nuchal band and
4–5 dark wide transverse bands with smooth edges.
Widths of interspaces between dorsal bands are less
than widths of the bands. Dorsal surface of head is
light yellowish, without any clear pattern. Dark
wide transverse bands present on the tail. Roundish
weakly keeled tubercles present on the dorsum, hind
limbs, temporal region of the head, on the tail base
and absent on the dorsal surface of the head. Ventrals
in 32–38 longitudinal rows at midbody. Lateral folds
weakly developed. Continuous series of 40–44 precloacal and femoral pores in males reach the bend of
knee. Enlarged femoral scales present, 5–6 enlarged
postcloacal spurs in both sexes. Subcaudal scales
transversally enlarged in one median row.
Holotype description. Cyrtodactylus khammouanensis sp. nov. – adult male, the holotypehas the following measurements: SVL 70.8 mm, TailL 95.0 mm,
HeadL 19.8 mm, HeadW 12.2 mm, HeadH 7.4 mm ,
SnEye 7.5 mm, OrbD 4.4 mm, EarL 2.0 mm, EyeEar
5.2 mm; proportions are as follows: SVL/HeadL 3.57,
HeadL/HeadW 1.62, HeadL/HeadH 2.67, SnEye/
EyeEar 1.44.
A review of the genus Cyrtodactylus
407
Fig. 4. Cyrtodactylus khammouanensis sp. nov.: A – general view of type specimen; B – precloacal region with continuous row of femoral
and precloacal pores; C – ventral surface of the head with mental scalation; D – dorsal surface of the head without enlarged tubercles;
E – dorsum surface with the rows of enlarged tubercles.
408
Rostral large, somewhat wider than high (RW
3.0 mm, RH 1.9 mm, RW/RH 1.57) with an inverse
Y-shaped median suture; supralabials 12/12; infralabials 10/11; nares surrounded by rostral anteriorly,
first supralabial laterally, supranasal and two nasals
posteriorly; rostral about 7–8 times larger than
supranasal; supranasals in contact with each other;
dorsum of head covered by small granular scales
without enlarged keeled tubercles (Fig. 4c); on the
temporal region rounded, weakly keeled tubercles,
which are ca. three times larger than the surrounding scales, are present; mental pentagonal, as wide as
rostral; two pairs of enlarged postmentals, first pair
longer than wide in broad contact (Fig. 4d); dorsal
scales granular, 2–3 times smaller than the ventral
scales; dorsal tubercles not large (3–4 times larger
than the surrounding scales), conical, weakly keeled,
surrounded by 8–10 granular scales, tubercles forming about 18 irregular longitudinal rows at midbody
(Fig. 4e); ventral scales smooth, 38 longitudinal rows
at midbody; lateral folds weakly developed; dorsal
surface of forelimbs without enlarged tubercles and
hindlimbs with smooth roundish enlarged tubercles;
fingers and toes without web, basal lamellae more
rounded than on distal surface of digits; row of enlarged precloacal and femoral scales with 44 pores
(Fig. 4b); five pairs of enlarged postcloacal spurs;
dorsal surface of tail without enlarged keeled tubercles; posterior part of tail covered by flattened and
rounded scales; subcaudals flat, smooth, imbricate,
with enlarged median plate row, about two times
wider than high.
Coloration: Dorsal head surface light without any
markings; nuchal band broadened posteriorly, dark
brown with distinct white margin, extending from the
neck to the posterior edges of eyes; labials light with
brown spots. Dorsum brownish with six wide regular,
dark, transverse bands with light posterior margins;
flanks grayish white. Venter white, the lower side of
toes and fingers light colored; dorsal surface of limbs
and digits brownish without any contrasting markings. Dorsal surface of tail with 3 dark brown bands
which are wider than the light bands in-between.
Ventral sides of tail dark grey. For coloration in life
see Fig. 4a.
Variation of paratypes. For the variation of the
type series see Table 4. The dorsal pattern is nt variable. Sexual dimorphism is pronounced, precloacal
and femoral pores present in males only; males also
have better developed postcloacal spurs.
R.A. Nazarov et al.
Comparisons with Laotian congeners. Cyrtodactylus khammouanensis sp. nov. differs from C. buchardi by the singe median row of subcaudals (vs.
subcaudals not enlarged in C. buchardi), enlarged
femoral scales (lacking in C. buchardi), fewer dorsal
tubercle rows (16 or 21 vs. 25), greater number of
ventral scales (32–38 vs. 30), and greater number
of subdigital lamellae under the fourth finger and
toe (18 or 20 and 20–23 vs. 14 and 12, respectively),
moreover, dorsal pattern of the new species consists
of six wide regular, dark, transverse bands versus five
transverse series of irregular blotches in C. buchardi.
From C. interdigitalis the new species clearly differs
by transverse enlarged subcaudal scales; higher number of precloacal and femoral pores in the singe row
(40–44 in the new species vs. 14 precloacal and 9+9
femoral pores in C. interdigitalis); roundish tail versus
flattened tail; dorsal pattern of the new species consists of six wide regular, dark, transverse bands versus
4–5 wide brownish transverse bands with jagged edges; no webbing between toes versus developed web on
the basis of toes. Cyrtodactylus khammouanensis sp.
nov. seems to be closely related to recently described
C. jaegeri and differs from this species by the following combination of morphological attributes: the new
species has small smooth and round dorsal tubercles
in 16–21 irregular longitudinal rows at midbody,
which are not present on the occipital region versus
round conical dorsal tubercles in 15–17 irregular
longitudinal rows which continue on the occipital
region, the new species has yellowish head coloration
with a wide nuchal loop (approximately the same size
as transverse bands, located posteriorly) versus light
brown head with narrow nuchal loop (notably more
narrow than the subsequent transverse dorsal bands),
somewhat greater number of ventral scales (32–38 vs.
31–32). These two species can be further diagnosed
by body and head proportions: the new species has
relatively smaller head (SVL/HL 3.57 vs. 3.27); comparatively smaller orbit (HL/OrbD 4.5 vs. 3.89) and
comparatively larger temporal region (HL/EyeEar
3.8 vs. 4.81). The new species differs from C. jarujini
by the smaller body size (73 mm vs. 90 mm in C. jarujini), lesser number of femoral and precloacal pores
(40–44 vs. 52–54), dorsal color pattern (banded vs.
blotched in C. jarujini). The new species is similar to
C. lomyenensis but is distinguishable from it by the
following combination of morphological characters:
lesser number of dorsal tubercle rows (16–21 vs. 20–
24), continuous row of femoral and precloacal pores
A review of the genus Cyrtodactylus
409
Table 4. Measurements and selected morphological characters of the type series of Cyrtodactylus khammouanensis sp. nov.; f = female,
m= male.
Holotype
Paratypes
ZMMU R-13982
ZIN 28250
ZMMU R-13983
ZIN 28251
Sex
m
m
f
f
68.5
SVL
70.8
73
72
TailL
95
83
90*
*
Head L
19.8
20.0
19.2
19.3
Head W
12.2
12.2
12
12.5
Head H
7.4
7.6
7.5
8.0
SnEye
7.5
7.8
7.7
7.2
OrbD
4.4
4.8
4.8
4.3
EarL
2.0
2.1
1.8
2.0
EyeEar
5.2
4.7
4.7
4.8
TrunkL
30.7
30.8
32
29
LS
11.6
11.6
11
11.8
ForeaL
10.8
10.7
11
10.5
FemurL
16.5
14.8
15.2
15
Crus L
12.7
12.3
12
11.7
LD4A
6.6
6.2
7.2
6.4
LD4P
8.7
9.3
8.2
8.6
V
38
32
34
36
155
SLB
169
172
159
LF 4
18
19
20
19
LT 4
20
22
22
23
PP+FP
44
40
0
17
SL
12
11
11
11
IL
10
10
10
9
TubL
37
48
47
45
TubW
18
16
21
19
SLH
87
83
87
77
SAH
63
66
57
56
reaching the bend of knee versus not reaching at one
third in C. lomyenensis, and dorsal pattern (bands
with smooth margins vs. bands with wavy margins);
furthermore, the new species has relatively smaller
orbit (HL/OrbD 4.5 vs. 3.75) and relatively larger
temporal region (HL/EyeEar 3.8 vs. 3.3). Cyrtodactylus khammouanensis sp. n. differs from C. pageli
by the following characters: continuous row of precloacal and femoral pores (40–44 vs. 4–6 precloacal
pores), lesser number of ventral scale rows (32–38 vs.
41–46), greater number of enlarged dorsal tubercle
rows (16–21 vs. 9–14), and the dorsal pattern (bands
with smooth margin vs. bands with wavy margins in
C. pageli). The new species is distinguishable from C.
roesleri by having a greater number of precloacal and
femoral pores (40–44 vs. 20–28) and by dorsal pattern (the width of dorsal transverse bands is greater
than the widths of interspaces between them vs. the
width of dorsal bands is less than of the interspaces
in-between). The new species differs from C. teyniei
by smaller body size (maximum SVL 73 mm vs. 89.9
mm in C. teyniei), dorsal pattern (bands vs. blotched
pattern in C. teyniei), nuchal loop (present vs. absent
in C. teyniei). The new species is distinguished from
410
C. puhuensis by having greater number of precloacal
and femoral pores (40–44 vs. 5 precloacal pores in
C. puhuensis), and by a different dorsal color pattern
(dark broad bands vs. light narrow bands).
From C. vilaphongi, the new species can be distinguished by having an enlarged median row of
subcaudals and a different dorsal color pattern (dark
broad bands vs. narrow yellowish white bands in
C. vilaphongi). From C. wayakonei the new species
is distinguished by having a greater number of precloacal and femoral pores (40–44 vs. 6–8 precloacal
pores), by having a different dorsal color pattern
(dark broad bands on the dorsum and no patterns
on the head vs. head and dorsum with blotched to
reticulated pattern in C. vilaphongi), and by differences in subcaudal scalation (distinctly enlarged
median row of subcaudals vs. somewhat enlarged
and broadened subcaudals in C. vilaphongi). Cyrtodactylus khammouanensis sp. nov. differs from
Cyrtodactylus darevskii sp. nov. by smaller body size
(maximum SVL 73 mm vs. 100 mm in C. darevskii
sp. nov.), fewer numbers of ventral scales (32–38 vs.
38–46) and dorsal color pattern (wide transverse
bands vs. narrow transverse bands).
Etymology. The specific epithet of the new
species is derived from the name of Khammouane
Province of Laos, renowned for its great diversity of
Cyrtodactylus geckoes. Suggested common name in
English: Khammouane Bent-toed Gecko.
Habitat description. The new species was recorded in the environs of Na Phao Village, Boulapha District, Khammouane Province, eastern Laos.
(17°34´57.1´´N and 105°44´37.3´´E, elevation 170 m
a.s.l.). Field work was conducted in this area during
15–20 June, 2009. The study area is situated near
Phou Hinboun (Limestone) National Biodiversity
Conservation Protected Area. The area is dominated
by sparsely vegetated limestone karst. Cyrtodactylus
khammouanensis sp. nov. was found at the altitude
170 m a.s.l. in a karst cave (Fig. 8). All specimens
were found on the walls of the cave; the new species
shares this habitat with Cyrtodactylus darevskii sp.
nov.
Distribution. The new species is to date known
only from the envrions of the type locality.
Phylogenetic position. A member of C. phongnhakebangensis species group; most closely related to
C. roesleri, distributed in Quang Binh Province of
Vietnam and Khammouane Province of Laos (p =
16.42–16.69%).
R.A. Nazarov et al.
Cyrtodactylus multiporus sp. nov.
(Fig. 5)
Holotype. Adult male ZMMU R–13984 collected
on 19 June 2009 by Nikolai Orlov, Sang Ngoc Nguyen
and Konstantin Milto in the environs of Na Home
Village, Boulapha District, Khammouane Province,
eastern Laos (17°32´40.3´´N and 105°41´43.0´´E,
elevation 230 m a.s.l.).
Paratypes. Two adult males ZMMU R–13985–1
(FN 4); ZIN 28252 (FN 3), six adult females ZMMU
R-13985-2; 13985-3; 13985-4 (FN 5, 6, 7); ZIN
28253; 28254; 28255 (FN 1, 2, the third without
field number), and three subadult specimens ZMMU
R-13985-5 (FN 8); ZIN 28256; 28257 (FN 9, 10). All
members of the type series have the same collection
data as the holotype.
Diagnosis. Relatively large-sized species of
Cyrtodactylus with a maximum SVL of 98 mm; the
new species is distinguished from its congeners by a
combination of the following morphological features.
Dorsal pattern consisting of dark irregular separate
spots and blotches between limbs and on the dorsal
part of the head. The nuchal band is not developed,
6–8 dark wide transversal bands with irregular edges
present on the dorsal surface of tail. Small smooth
roundish dorsal tubercles present on the occipital
region and sides of the head, body, hind limbs and the
base of tail. Ventrals in 30–38 longitudinal rows at
midbody. Continuous series of 58–60 precloacal and
femoral pores in males, 5–6 postcloacal spurs in both
sexes. Tail not segmented without whorls and keeled
tubercles on its dorsal surface, one median row of
transversally enlarged subcaudal scales.
Holotype description. Adult male, medium sized;
the holotype has the following measurements: SVL
86.8 mm, TailL 95.0 mm, HeadL 24.2 mm, HeadW
15.3 mm, HeadH 9.0 mm , SnEye 9.6 mm, OrbD
5.8 mm, EarL 2.4 mm, EyeEar 5.0 mm; proportions
are as follows: SVL/HeadL 3.58, HeadL/HeadW
1.58, HeadL/HeadH 2.68, SnEye/EyeEar 1.92. Rostral is large, somewhat wider than high (RW 4.0 mm,
RH 2.25 mm, RW/RH 1.7) with an inverse T-shaped
suture on the median part; supralabials 10/10; small
scales between orbit and the seventh supralabial
4/4; infralabials 11/9; nares surrounded by rostral
anteriorly, first supralabial laterally, supranasal and
3 nasals posteriorly; rostral about 7–8 times larger
than supranasal; supranasals in contact to each other;
upper anterior ciliaries two times larger than poste-
A review of the genus Cyrtodactylus
411
Fig. 5. Cyrtodactylus multiporus sp. nov.: A – general view of type specimen; B – precloacal region with continuous row of femoral and
precloacal pores; C – ventral surface of the head with mental scalation; D – dorsal surface of the head with enlarged tubercles; e – dorsum
surface with the rows of enlarged tubercles.
412
rior ones; head scales granular, some smaller than the
median snout scales; dorsum of head and temporal
region with rounded, keeled tubercles, that are 2–3
times larger than the surrounding scales (Fig. 5c);
mental triangular; one pair of enlarged postmental,
longer than wide, in broad contact (Fig. 5d); dorsal
scales granular, 3–4 times smaller than the ventral
scales; dorsal tubercles round, flat, not keeled, surrounded by 7–10 granular scales, tubercles forming
about 20 irregular longitudinal rows at midbody
(Fig. 5e); ventral scales smooth, 30 longitudinal rows
at midbody; lateral folds weakly developed; dorsal
surface of fore- and hindlimbs with granular scales
and weakly keeled conical tubercles; fingers and toes
without web, basal lamellae more rounded than on
distal surface of digits; a row of precloacal scales with
58 pores (Fig. 5b); enlarged femoral scales (without
pores) are present; five pairs of enlarged postcloacal
spurs; not segmented tail without whorls and keeled
tubercles on dorsal surface; subcaudals with row of
enlarged plates, smooth and imbricate.
Coloration: Dorsal surface of head is light brown
with small irregular rounded dark brown spots; continuous nuchal band not developed; labials brownish
grey with white spots. Dorsal body surfaces brownish
with dark irregular separate spots with light margins.
Ventral surface is white, the lower side of toes and fingers grey; dorsal surface of limbs and digits brownish
with irregular bands. First third of the tail with three
dark brown bands; they are wider than the light-grey
interspaces in-between them. Ventral side of tail dark
brown with light spots that are becoming more distinct posteriorly. For coloration in life see Fig. 5a.
Variation of paratypes. For the variation of the
type series see Table 5. The dorsal pattern is somewhat variable. Sexual dimorphism is well developed,
males are smaller then females, maximal SVL for
males is 86.8 mm and for females 98 mm; precloacal
pores are present in males only and males have also
more developed postcloacal spurs.
Comparisons with Laotian congeners. Cyrtodactylus multiporus sp. nov. differs from C. buchardi by
the singe median row of subcaudals (vs. subcaudals
not enlarged in C. buchardi), enlarged femoral scales
(lacking in C. buchardi), fewer dorsal tubercle rows
(16–20 vs. 25), more ventral scales (30–38 vs. 30),
and more subdigital lamellae under the fourth finger
and toe (18–20 and 18–22 vs. 14 and 12, respectively).
From C. interdigitalis the new species is clearly different in having transversal enlarged subcaudal scales;
R.A. Nazarov et al.
higher number of precloacal and femoral pores in the
singe row (58–60 in the new species vs. 32 precloacal
and femoral pores in C. interdigitalis); roundish tail
versus flattened tail; blotched irregular dorsal pattern
versus 4–5 wide brownish jagged transversal bands;
no webbing between toes versus developed webbing
on the toe basis in C. interdigitalis.
Cyrtodactylus multiporus sp. nov. differs from
C. jaegeri by larger body size (maximum SVL 98 mm
vs. 68.5 mm); blotched dorsal patterns versus wide
transversal bands; small, smooth and rounded dorsal
tubercles in 16–20 longitudinal rows at midbody
versus round conical dorsal tubercles in 15–17 longitudinal rows; in lacking a continuous nuchal loop
versus distinct nuchal loop in C. jaegeri; somewhat
greater number of ventral scales (30–38 vs. 31–32).
The new species is similar to C. jarujini by the
body size and dorsal color pattern but can be diagnosed from this species by having a higher number
of femoral and precloacal pores (58–60 pores in continuous row vs. 52–54 pores in irregular rows) and
in having more subdigital lamellae (LF4 18–20 and
LT4 18–22 vs. 11–17 and 11–18 respectively). From
C. lomyenensis the new species differs by the larger
body size (maximum SVL 98 mm vs. 72.1 mm in
C. lomyenensis), a higher number of pores (58–60 vs.
32–40), lesser number of dorsal tubercle rows (16–20
vs. 20–24), and by dorsal color pattern (blotches vs.
wide bands in C. lomyenensis). Cyrtodactylus multiporus sp. nov. can be differentiated from C. pageli
by the following characters: a continuous row of
precloacal and femoral pores (58–60 vs. 4–6 precloacal pores in C. pageli), lesser number of ventral scale
rows (30–38 vs. 41–46), greater number of enlarged
dorsal tubercle rows (16–20 vs. 9–14) and dorsal
pattern (separate irregular spots vs. bands with wavy
margins). The new species is distinguishable from C.
roesleri by having a higher number of precloacal and
femoral pores (58–60 vs. 20–28), a higher number
of enlarged dorsal tubercle rows (16–20 vs. 10–12),
by different dorsal pattern (blotches vs. wide dorsal
bands in C. roesleri), by lacking nuchal band versus
distinct nuchal loop in C. roesleri. The new species
is morphologically similar to C. teyniei and can be
distinguished from this species by lacking of femoral
and precloacal pores in females versus 13 or 14 pores
in the single known female of C. teyniei; by numeorus
roundish dark spots on dorsal surface of head versus
few dark spots on the head; by the dorsal color pattern having wide roundish dark blotches (vs. dorsal
A review of the genus Cyrtodactylus
413
Table 5. Measurements and selected morphological characters of the type series of Cyrtodactylus multiporus sp. nov.; f = female, m= male,
sub = subadult.
ZIN 28253
f
f
f
f
ZIN 28257
ZMMU R- 13985-4
m
ZIN 28256
ZMMU R- 13985-3
m
ZMMU R- 13985-5
ZMMU R- 13985-2
m
ZIN 28255
ZMMU R- 13985-1
Sex
ZIN 28254
ZIN 28252
Paratypes
ZMMU R-13984
Holotype
f
f
sub
sub
Sub
54
SVL
86.8
81.3
84.6
86.6
81
79.6
82
91
98
54.3
53.3
TailL
95*
85*
25*
105
98
97
102
100
76*
55
62
*
Head L
24.2
22.7
23.5
23.7
22.2
21.3
24
23.6
25.7
15.8
15.4
16.2
Head W
15.3
14.8
15.7
15.7
14.7
14.8
15.3
16.7
16.7
10
10.3
10.3
Head H
9.0
8.8
9.3
9.3
8.8
8.6
8.9
10
9.5
6.5
6.4
6.3
SnEye
9.6
9.1
9.3
9.4
8.8
8.4
8.8
9.8
10
6.3
6.2
6.3
OrbD
5.8
5.0
5.2
5.6
4.5
4.7
5.1
5.2
5.5
2.8
3.5
3.4
EarL
2.4
2.2
2.4
2.5
2.0
2.5
2.5
2.1
2.8
1.5
1.3
1.3
EyeEar
5.0
5.4
6.0
5.9
5.2
5.0
5.8
5.8
6.0
4.1
3.6
4.2
TrunkL
40.2
38.3
37.8
40.8
36.0
36
40.8
40.7
43.7
21.2
21.8
23.1
LS
15.2
14.5
14.1
14.5
12.9
13.2
13.8
14.7
15
9.4
8.6
9.2
ForeaL
13.2
12.8
13.2
13.5
12.5
12.0
12.7
13.6
13.3
8.7
8.0
8.2
FemurL
17.5
17.6
18.6
18.2
16.8
15
16.9
18.6
18.6
12.1
11.4
12.1
Crus L
14.1
13.6
14.2
14.8
13.0
12.8
13.5
14.0
14.4
9.0
9.4
9.6
LD4A
8.8
8.2
9.2
8.5
8.4
7.8
8.6
8.6
9.3
6.2
5.2
5.8
LD4P
11
9.5
9.6
10.4
9.5
10.0
9.6
9.8
10
6.7
6.6
6.3
V
30
30
36
37
36
34
36
38
34
34
37
32
SLB
174
168
168
178
173
177
180
171
181
177
164
174
LF 4
18
19
18
18
19
18
20
18
18
20
18
19
20
LT 4
22
19
22
21
19
18
20
19
18
22
19
PP+FP
58
60
58
0
0
0
0
0
0
0
0
0
SL
10
11
9
10
9
10
11
10
9
10
11
11
IL
11
11
10
10
9
9
11
10
9
10
10
10
TubL
42
44
42
41
41
34
45
43
42
45
41
40
TubW
20
18
20
20
18
16
18
18
18
18
18
18
SLH
89
93
96
94
88
81
96
89
86
92
89
100
SAH
89
76
72
79
75
68
83
78
76
73
70
80
pattern formed by thin and elongate blotches in
C. teyniei). The new species is distinguished from
C. puhuensis by having a higher number of precloacal and femoral pores (58–60 vs. 5 precloacal pores),
and a different dorsal pattern (dark blotches vs. light
narrow bands in C. puhuensis). From C. vilaphongi
the new species difers in having an enlarged median
row of subcaudals and a different dorsal color pattern
(dark blotches vs. narrow yellowish white bands in
C. vilaphongi). From C. wayakonei the new species is
distinguished by having a higher number of precloacal
and femoral pores (58–60 vs. 6–8 precloacal pores),
414
and different subcaudal scalation (distinctly enlarged
median row vs. subcaudals somewhat enlarged,
broadened in C. wayakonei). From Cyrtodactylus
darevskii sp. nov. the new species differs in lacking
of pores in females versus 24–34 pores in C. darevskii
sp. nov. females; and also in higher number of pores
in males (58–60 vs. 38–44), nuchal loop (absent vs.
present in C. darevskii sp. nov.). The new species is
distinguished from Cyrtodactylus khammouanensis
sp. nov. by its larger body size (maximum SVL 98 mm
vs. 73 mm in C. khammouanensis sp. nov.), and by
higher number of pores (58–60 vs. 40–44).
Etymology. The species epithet “multiporus” is
a Latin adjective in masculine, indicating the high
number of precloacal and femoral pores typical for
this species; derived from “multus” – “many” (Latin)
and “porus” – “pore”, “canal” (Latinized Greek).
Habitat description. The new species was recorded in a limestone forested area near Na Home Village
in Boulapha District, Khammouane Province, eastern
Laos (17°32´40.3´´N and 105°41´43.0´´E, elevation
230 m a.s.l.). Field work was conducted there during
15–19 June, 2009. The study area is situated near
Phou Hinboun (Limestone) National Biodiversity
Conservation Protected Area. The area is dominated
by sparsely vegetated limestone karst (Fig. 9).
Distribution. At the present moment the new
species is known only from the environs of the type
locality.
Phylogenetic position. A member of C. phongnhakebangensis species group; very closely related to
C. teyniei, also found in Khammouane Province of
Laos; uncorrected genetic distance between C. teyniei
and the new species comprise p = 6.00%.
Cyrtodactylus spelaeus sp. nov.
(Figs. 6–7)
Holotype. Adult male ZMMU R–14399 from
Khuang Lang Cave National Park, Kasi District, Vientiane Province, Laos (18°09.876´N and 104°30.387´E,
elevation 183 m a.s.l.). Collected on 22 November
2011 by E.L. Konstantinov and A.B. Gavrilov.
Paratypes. One adult female ZIN 28258 and one
subadult male ZMMU R–14400. All specimens of
the type series have the same collection data as the
holotype.
Diagnosis. The new species of Cyrtodactylus with
a maximum SVL of 91 mm, is distinguished from
all other congeners by the following combination of
R.A. Nazarov et al.
morphological characteristics. Dorsal pattern formed
by oblong dark irregular butterfly-shaped blotches
(Fig. 6a), with light contrast margins and a light middorsal medial stripe running across them. Nuchal
band with light edging consists of two separate parts,
which contact in the occipital region. Dorsal head
surface is light brown with roundish dark spots. The
intact (not regenerated) tail with ten dark transverse
bands. Roundish weakly keeled tubercles present on
the dorsum, limbs, temporal region of the head and
absent on the dorsal surface of the head. Ventrals in
36–39 longitudinal rows at midbody. Lateral folds
weakly developed. Males with 8–9 precloacal pores
in an angular continuous series. No femoral pores
and enlarged femoral scales, 2–3 enlarged postcloacal spurs. One median row of transversally enlarged
subcaudals.
Holotype description. The male of Cyrtodactylus
spelaeus sp. nov. have the following measurements:
SVL 88.9 mm, regenerated tail 80.1 mm, HeadL
26.3 mm, HeadW 16.9 mm, HeadH 10.8 mm , SnEye 10.2 mm, OrbD 5.6 mm, EarL 2.3 mm, EyeEar
8.0 mm; proportions are as follows: SVL/HeadL 3.38,
HeadL/HeadW 1.55, HeadL/HeadH 2.43, SnEye/
EyeEar 1.27.
Rostral large, somewhat wider than high (RW
3.9 mm, RH 2.5 mm, RW/RH 1.56) with a median
groove; supralabials 12/11; infralabials 9/10; nares
surrounded by rostral anteriorly, first supralabial laterally, supranasal and two nasals posteriorly; rostral
about 5–6 times larger than supranasal; supranasals
separated by one small scale from each other; dorsal
surface of the head covered by small granular scales
without enlarged tubercles (Fig. 7a); on the temporal
region rounded, weakly keeled tubercles present;
mental triangular, as wide as rostral; two pairs of enlarged postmentals, first pair in broad contact (Fig.
7b); dorsal scales granular, 2–3 times smaller than
the ventral scales; dorsal tubercles smooth, rounded,
not large (5–6 times larger than surrounding scales),
surrounded by 8–9 granular scales; tubercles form
ten irregular longitudinal rows at midbody (Fig. 7c);
ventral scales smooth, 39 longitudinal rows at midbody; lateral folds weakly developed; dorsal surface
of fore- and hindlimbs covered by smooth rounded
enlarged tubercles; no webbing on fingers and toes,
basal lamellae more rounded than on the distal part of
digits; eight precloacal pores in an angle series (Fig.
6b); enlarged postcloacal spurs – three (on the right
side) and two (on the left side); dorsal surfaces of the
A review of the genus Cyrtodactylus
415
Fig. 6. Cyrtodactylus spelaeus sp. nov.: A – general view of type specimen; B – precloacal region with angular row of precloacal pores.
416
R.A. Nazarov et al.
Fig. 7. Morphological comparison of Cyrtodactylus spelaeus sp. nov. (A–C) and Cyrtodactylus wayakonei (D–F): A – dorsal surface of the
head; B – ventral surface of the head with mental scalation; C – dorsum surface with ten rows of enlarged tubercles; D – dorsal surface of
the head with reticulated pattern; E – ventral surface of the head with mental scalation; F – dorsum surface with higher number rows of
enlarged tubercles.
tail without enlarged keeled tubercles; posterior part
of regenerated tail covered by flattened and rounded
scales; subcaudals forming enlarged median row.
Coloration: Dorsal head surface light grey with
few irregular dark brown butterfly–shaped blotches;
nuchal band dark brown with light contrast margin,
consists of two separate parts which contact posteriorly, extending from the neck to the posterior corners
of eyes; labials light brown. Dorsum light with seven
irregular, dark brown transverse blotches with light
A review of the genus Cyrtodactylus
417
Table 6. Measurements and selected morphological characters of the type series of Cyrtodactylus spelaeus sp. nov.; f = female, m= male,
sub = subadult.
Holotype
Sex
Paratypes
ZMMU R – 14399
ZMMU R – 14400
ZIN 28258
m
f
m, sub.
SVL
88.9
91
61.8
TailL
80.1*
83*
69.3
Head L
26.3
26.4
19.6
Head W
16.9
18
12.1
Head H
10.8
10.4
7.5
SnEye
10.2
10.4
7.6
OrbD
5.6
6.1
4.6
EarL
2.3
2.3
1.5
EyeEar
8
7.6
5.5
TrunkL
30.8
42.8
23.8
LS
17.3
17
12.1
ForeaL
15.3
15.7
10.8
FemurL
20
21.4
15.1
Crus L
16.6
18.8
12.8
6.8
LD4A
9.1
10
LD4P
12.3
11.5
9
V
39
36
37
183
SLB
156
178
LF 4
19
20
19
LT 4
24
22
23
PP
8
0
9
SL
12
9
11
IL
10
8
9
TubL
24
28
32
TubW
10
10
10
SLH
90
92
89
SAH
84
82
81
margins, a median longitudinal light stripe runs
along the vertebral column flanked by dark blotches.
Flanks grayish white. Venter white, the lower side
of toes and fingers light; dorsal surface of limbs and
digits light gray with brown transverse bands. Tail
basis with three dark brown bands which are wider
than the light interspaces in-between, bands absent
on regenerated part of the tail. Ventral surface of tail
light. For coloration in life see Fig. 6a.
Variation of paratypes. For the variation of the
type series see Table 6. Sexual dimorphism is well
developed, precloacal and femoral pores are present
in males only and males have better developed postcloacal spurs.
Comparisons with Laotian congeners. The new
species Cyrtodactylus spelaeus sp. nov. differs from
C. buchardi by having the singe median row of subcaudals and enlarged femoral scales (both lacking in
C. buchardi), fewer dorsal tubercle rows (10 vs. 25),
more ventral scales (37–39 vs. 30), and greater number of subdigital lamellae under the fourth finger and
toe (19–20 and 22–24 vs. 14 and 12, respectively).
The new species can be distinguished from C. interdigitalis by having transversal enlarged subcaudal
418
R.A. Nazarov et al.
Fig. 8. Habitat of Cyrtodactylus darevskii sp. nov. and Cyrtodactylus khammouanensis sp. nov. nearly Na Phao Village, Boulapha District,
Khammouane Province, Laos.
scales (absent in C. interdigitalis); lesser number of
precloacal and femoral pores in the singe row (8–9
vs. 32 in C. interdigitalis); roundish tail versus flattened tail; blotched dorsal pattern in the new species
versus 4–5 wide brownish jagged transversal bands;
no webbing between toes versus developed web on
toe basis in C. interdigitalis. Cyrtodactylus spelaeus
sp. nov. can be diagnosed from C. jaegeri by the larger body size (maximum SVL 91 mm vs. 68.5 mm), a
higher number of ventral scales (37–39 vs. 31–32)
and dorsal color pattern (dark irregular butterflyshaped blotches vs. wide transversal dark bands in C.
jaegeri). The new species differs from C. jarujini by
having lesser number of pores (8–9 vs. 52–54 in C.
jarujini), more ventral scales (37–39 vs. 30–38), and
greater number of subdigital lamellae (LF4 19–20
and LT4 23–24 vs. 12–17 and 11–18 respectively).
The new species is distinguishable from C. lomyenensis by its larger body size (maximum SVL 91
mm vs. 72.1 mm in C. lomyenensis), a higher number
of ventral scales (37–39 vs. 35–36), and dorsal coloration pattern (butterfly-shaped blotches vs. wide
bands in C. lomyenensis). Cyrtodactylus spelaeus sp.
nov. differs from C. pageli by the following morphological characters: dorsal color pattern (blotches vs.
transversal dark bands in C. pageli), greater number of precloacal pores (8–9 vs. 4–6 in C. pageli),
somewhat fewer rows of enlarged dorsal tubercles
(10 vs. 9–14 in C. pageli). From C. roesleri the new
species is distinguishable by having a lesser number
of pores (8–9 vs. 20–28), larger maximum body size
(91 mm vs. 75.3 mm) and by dorsal pattern (irregular, dark blotches vs. wide transverse dark bands in
C. roesleri). The new species differs from C. teyniei
in absence of pores in females versus 13 or 14 pores
known for C. teyniei, as well as in dorsal coloration
pattern (dark irregular butterfly-shaped blotches vs.
dark brown oblong blotches in C. teyniei) and presence of a nuchal loop (vs. nuchal loop absent in C.
teyniei).
A review of the genus Cyrtodactylus
419
Fig. 9. Habitat of Cyrtodactylus multiporus sp. nov. in environs of Na Home Village, Boulapha District, Khammouane Province, Laos.
The new species is distinguishable from C. puhuensis by a higher number of precloacal pores (8–9 vs. 5
precloacal pores in C. puhuensis), and by a different
dorsal coloration pattern (dark irregular blotches vs.
light narrow bands in C. puhuensis). The new species
differs from C. vilaphongi in having enlarged median
row of subcaudals, higher number of ventral scales
(37–39 vs. 34–36) and different dorsal coloration
pattern (irregular, dark blotches vs. narrow yellowish
white bands in C. vilaphongi).
The new species is morphologically quite similar
with C. wayakonei, sharing many diagnostic features,
such as the number of precloacal pores 8–9 in the new
species versus 6–8 in C. wayakonei, quite similar dorsal color pattern, lacking of femoral pores and similar
state of subcaudal scalation in both species. But the
new species is greately different from C. wayakonei in
COI gene sequences (p = 16.7%) and can be further
diagnosed from this species by having a fewer number
of longitudinal rows of enlarged dorsal tubercles (10
vs. 17–19 in C. wayakonei) (Fig. 7c,f), somewhat
higher number of subdigital lamellae (LF4 19–20
and LT4 22–24 vs. 17–18 and 19–20 respectively),
by presence of distinct nuchal band versus not developed nuchal band in C. wayakonei, and by dorsal
surface of head having few dark roundish spots versus
reticulated grey-brown patterns in C. wayakonei
(Fig. 7a, d).
Etymology. The species epithet “spelaeus” is a
Latin adjective in masculine, derived from the Latin
“spelaeum”, “spelaean”, “living in a cave” referring
to the habitat and the type locality of the new species – the Khuang Lang Cave in Kasi District of the
Vientiane Province of Laos.
Habitat description. The new species inhabits
typical limestone karst area covered with lush vegetation. All specimens were found on the wall inside
the karst cave.
Distribution. To date the new species has been
reported only from the environs of its type locality
420
in Khuang Lang Cave National Park, Kasi, Vientiane
Province, Laos.
Phylogenetic position. A member of C. wayakonei species group; most closely related to a clade
joining C. vilaphongi from Luang Prabang Province
of Laos and C. puhuensis from Houphan Province of
northern Laos and Thanh Hoa Province of northern
Vietnam; uncorrected genetic distance between these
taxa and the new species comprise p = 11.48–12.30%.
DISCUSSION
Taxonomic diversity of South-East Asian benttoed geckoes is astonishing and in many areas still
remains unexplored. In Vietnam, the number of
described species of the genus Cyrtodactylus has
increased rapidly since 2007, growing from 5 species known in 2006 to 33 species recognized in 2014
(Nguyen et al. 2014). However diversity of Cyrtodactylus in such areas of Indochina as Laos and Cambodia is comparatively poorly studied (Schneider et al.
2014b). The recent and rapidly increasing number of
described species indicates the need for further exploration. DNA barcoding (using fragment sequences
of COI, cytochrome c oxidase subunit I) allows the
estimation of biodiversity and species identity, and
serves an efficient tool may point to populations in
need of further investigation and was successfully applied to many groups of reptiles, including the genus
Cyrtodactylus (Nazarov et al. 2012; Nagy et al. 2012;
Murphy et al. 2013; Nguyen et al. 2013, 2014; Schneider et al. 2014a, 2014b).
Herein we successfully applied COI DNA barcoding for scrutinizing diversity of Laotian Cyrtodactylus. This work includes 11 of 15 recognized species
(73.3%) of Cyrtodactylus in Laos, including our four
new species. At least one additional undescribed
taxon occurs in Luang Prabang Province of Laos
(Schneider et al. 2014b). DNA barcoding using COI
efficiently guided species delimitation and discovery
of new lineages (Nazarov et al. 2012; Nguyen et al.
2014). Further application of molecular methods
including COI DNA barcoding is required for proper
assessment of Cyrtodactylus taxonomic diversity.
Application of molecular methods is also important for re-evaluation of morpohological characters
used in Cyrtodactylus taxonomy. Such important features of external morphology as subcaudal scalation,
number and position of precloacal and femoral pores
R.A. Nazarov et al.
agree well with grouping of taxa proposed by the
results of COI barcoding. Thus, the tree species of
Cyrtodactylus described from Phong Nha – Ke Bang
National Park in Quang Binh Province of Vietnam
(C. phonghnakebangensis, C. roesleri and C. cryptus)
are superficially quite similar to each other and have
similar dorsal patterns. However, C. cryptus (showing absence of enlarged subcaudals and femoral
pores; features characteristic to the members of C.
irregularis species complex), based on the phylogenetic hypothesis derived from COI sequence data
and in full agreement with morphology, is clearly
grouped with other members of C. irregularis species
complex.
Morphologically, Cyrtodactylus darevskii sp. nov.
is most similar to C. teyniei from Khammouane Province, especially in its dorsal pattern. Both species
are limestone-dwelling bent-toed geckos and the
distance between the type localities of these species
is approximately 100 km. Nonetheless, C. darevskii
sp. nov. is distantly related to C. teyniei based on
the molecular data, which suggests that similarity
in dorsal pattern and other morphological features
might be caused by morphological convergence evolution. Our analysis indicate that C. darevskii sp. nov.
is closely related to C. phongnhakebangensis, whereas
C. teyniei is genetically quite close to morphologically distant C. multiporus sp. nov. It is noteworthy
that both C. darevskii sp. nov. and C. phongnhakebangensis show well-pronounced comlete nuchal
loop, whereas both C. teyniei and C. multiporus lack
nuchal loop.
Three of the four new Cyrtodactylus species
described herein (C. darevskii sp. nov., C. khammouaensis sp. nov. and C. multiporus sp. nov.) were
found in the same locality in the environs of the Na
Home Village, Khammouane Province; two of them
(C. darevskii sp. nov., C. khammouaensis sp. nov.) occurred in the same biotope. This example illustrates
the amazing diversity of the genus Cyrtodactylus even
in a very limited area; it is undoubtful that number of
recognized Cyrtodactylus species will only continue
to grow in future.
According to our data, to date the following species of Cyrtodactylus are documented to occur in Laos:
C. buchardi, C. darevskii sp. nov., C. interdigitalis, C.
spelaeus sp. nov., C. jaegeri, C. jarujini, C. khammouanensis sp. nov., C. lomyenensis, C. multiporus sp. nov.,
C. pageli, C. puhuensis, C. roesleri, C. teyniei, C. vilaphongi, C. wayakonei.
A review of the genus Cyrtodactylus
ACKNOWLEDGEMENTS
Collecting permits were made available through National University of Laos, Faculty of Forestry Science,
Vientiane, Lao PDR and Agriculture and Forestry of
Khammouane Province, Lao PDR. Export permits were
made available through Division of Forest Resource
Conservation of Ministry of Agriculture and Forestry
Department of Forestry, Lao PDR. Special thanks to Head
of Wildlife Managements Section, Chaynoy Sisomphane,
Head of CITES Section, Bounsou Sovan and Deputy Head
of Forestry section, Khammoune Province, Soukanh Inthanouhack. We are indebted to all the people of Ban Doy
Community, Thakhek District, Khammouane Province,
Lao PDR for their invaluable help and cordial hospitality. We sincerely thank Evgeniya N. Solovyeva for help in
the laboratory and data analysis. This research was partly
supported by Joint Russian-Vietnamese Tropical Research
and Technological Center (JRVTRTC). Molecular analysis was partially carried out within the NPRPV project of
the BOLD. We thank Alex Borisenko, Natalia Ivanova and
Paul D.N. Hebert for collaboration. The research was partially supported by NSERC, Genome Canada, Gordon and
Betty Moore Foundation, Canada Foundation for Innovation and Ontario Innovation Trust, Canadian Centre for
DNA Barcoding (BIO, ON, Canada) and funded through
Genome Canada and the Ontario Genomics Institute
(2008-OGI-ICI-03). We thank Thomas Ziegler (Cologne
Zoo, Germany) for the loan of specimens and tissue samples
and his kind support and Le Duc Minh (Vietnam National
University, Vietnam) for providing sequences for analysis
and useful information. For permission to study specimens
under their care, we thank Valentina F. Orlova (ZMMU)
and Natalia B. Ananjeva (ZIN). The study was supported
by the Russian Foundation of Basic Research (Grants No.
RFBR_Taiwan No. 14-04-92000, RFBR 12-04-01552_a,
15-04-08393_а and 15-04-02029_a) and by Grants of the
President of Russian Federation (MK-5815.2014.4 and
NS-2990.2014.4).
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Submitted December 11, 2014; accepted December 20, 2014.