Research Article |
Corresponding author: Shauri Sulakhe ( shaurisulakhe@gmail.com ) Academic editor: Alexander Haas
© 2021 Amit Sayyed, Vivek Philip Cyriac, Anish Pardeshi, Shauri Sulakhe.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Sayyed A, Cyriac VP, Pardeshi A, Sulakhe S (2021) Dwarfs of the fortress: A new cryptic species of dwarf gecko of the genus Cnemaspis Strauch, 1887 (Squamata, Gekkonidae) from Rajgad fort in the northern Western Ghats of Maharashtra, India. Evolutionary Systematics 5(1): 25-38. https://doi.org/10.3897/evolsyst.5.62929
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A new species of the genus Cnemaspis Strauch, 1887 is described from the Rajgad fort in the northern Western Ghats of Maharashtra, India. The new species, belonging to the flaviventralis clade, is one of the smallest known Indian Cnemaspis and can be distinguished from other congeners by its genetic distinctiveness and few key morphological characters. The new species can be diagnosed from all other Indian congeners by its small body size (SVL < 27 mm), the absence of conical and spine-like tubercles on flank; heterogeneous dorsal pholidosis; presence of only femoral pores and no precloacal pores in males; weakly keeled scales on the ventral surface of neck, pectoral, abdominal region and limbs; granular scales on the tail with whorls of slightly enlarged, strongly keeled tubercles; and the absence of enlarged median subcaudal scales. The new species is currently known to inhabit the man-made historical structures from a single locality in Rajgad, Maharashtra, where it is presumed to be uncommon.
Cnemaspis rajgadensis sp. nov., taxonomy, dwarf gecko, cryptic species, reptiles, forts of Maharashtra
Western Ghats of India is a known global biodiversity hotspot with high levels of endemism (
The genus Cnemaspis Strauch, 1887 is one of the most diverse clades of geckos in India and recent studies have indicated that the diversity documented within this clade is far from complete (
The NWG in Maharashtrtra, popularly known as Sanhyadri, has been a region of strategic importance in several historical events due to its unique climatic conditions and geographic features. Many empires have occupied and ruled Maharashtra building more than 300 forts in NWG at vantage points on the lofty cliffs of the NWG, which are surrounded by dense forests (
Specimens were collected during September 2020, in parts of Rajgad Fort, Pune District, Maharashtra, India. A single adult male and two gravid females were collected by hand, photographed in life, and euthanized using halothane. Tail clips were collected as tissue sample and stored in absolute ethanol for genetic analysis, after which specimens were fixed in 4% formaldehyde for 24 hours, washed in water, and transferred to 70% ethanol for long-term storage. Scalation and other morphological characters were recorded using a Lensel stereo microscope. The materials referred to in this study are deposited in the collection of the Bombay Natural History Society (
DNA extraction, amplification and sequencing
Total genomic DNA was extracted from muscle tissue (tail clips) using a MACHEREY-NAGEL NucleoSpin DNA Insect kit following the manufacturer’s protocols. A 550–600 base pair (bp) fragment of the mitochondrial 16S rRNA gene was amplified by standard 3-step polymerase chain reaction (PCR) using published primers (
Sequence alignment
We added the newly generated sequences (see Table
Voucher numbers and GenBank accession numbers for the sequence data generated in this study and used in the phylogenetic analysis.
Species | Voucher Number | GenBank accession number | Locality |
---|---|---|---|
Cnemaspis girii | ASPC6 | MW682860 | Morewadi, Satara |
Cnemaspis girii | ASPC8 | MW682861 | Morewadi, Satara |
Cnemaspis sp. | ASPC11 | MW682862 | Amba Ghat, Kolhapur |
Cnemaspis sp. | ASPC12 | MW682863 | Amba Ghat, Kolhapur |
Cnemaspis koynaensis | ASPC10 | MW682864 | Humbarali, Koyana, Satara |
Cnemaspis koynaensis | ASPC9 | MW682865 | Humbarali, Koyana, Satara |
Cnemaspis rajgadensis sp. nov. |
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MW682866 | Rajgad fort, Pune |
Cnemaspis rajgadensis sp. nov. |
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MW682867 | Rajgad fort, Pune |
Molecular phylogenetic analysis
We removed ambiguously aligned positions from the 16S rRNA sequence matrix using GBlocks v.0.91b (
Morphological data were taken using a Yamayo digimatic calliper, a Mitutoyo 500, or a Tesacalip 64 to the nearest 0.1 mm. Morphological data recorded included the snout-vent length (SVL), distance from tip of snout to anterior margin of vent; trunk length (TRL), distance from axilla to groin; trunk width (TW), maximum width of the body; eye diameter (ED), horizontal diameter of the eye; eye-to-nares (EN), distance between anterior point of the eye to the posterior part of the nostril; snout length (ES), distance from anterior margin of the eye to the tip of the snout; eye-to-ear (ET), distance from posterior margin of the eye to the anterior margin of the ear opening; internarial distance (IN), least distance between the inner margins of the nostrils; ear opening diameter (EOD), horizontal distance from the anterior to posterior margin of the ear opening; head length (HL), distance from tip of snout to posterior edge of mandible; head width (HW), maximum width of the head; head depth (HD), maximum depth of the head; interorbital distance (IO), shortest distance between the superciliary scale rows; upper arm length (UAL), distance from axilla to elbow, lower arm length (FAL), distance from elbow to wrist; palm length (PAL), distance from wrist to the tip of the longest finger; finger length (FL), distance from the tip of the finger to the nearest fork; femur length (FEL), distance from groin to the knee; tibia length (TBL), distance from knee to heel toe length (TOL), distance from tip of 1st toe to the nearest fork; tail length (TL), distance between posterior margin of vent to the tip of the tail.
Meristic data recorded for all specimens included number of supralabials (SupL) and infralabials (InfL) on left (L) and right (R) sides; number of supraciliaries (SuS); number of interorbital scales (InO); number of scales between eye to tympanum (BeT), from posterior-most point of the orbit to anterior-most point of the tympanum; number of the postnasal (PoN), all scales posterior to the naris; number of postmentals (PoM);number of posterior postmentals (PoP), scales that are surrounded by the posterior-postmentals and between infralabials; number of supranasal (SuN), excluding the smaller scales between the larger supranasals; number of canthal scales (CaS), number of scales from posterior-most point of naris to anterior most point of the orbit; number of dorsal paravertebral scales (PvS), between pelvic and pectoral limb insertion points along a straight line immediately left of the vertebral column; number of mid-dorsal scales (MbS), from the centre of mid-dorsal row diagonally towards the ventral scales; number of mid-ventral scales (MvS), from the first scale posterior to the mental to last scale anterior to the vent; number of mid-body scales (BlS), across the ventral between the lowest rows of dorsal scales; femoral pores (FPores), the number of femoral pores; poreless scales (PS), number of poreless scales between left and right femoral pores; lamellae under digits of manus (MLam) and pes (PLam) on right (R) side, counted from first proximal enlarged scansor greater than twice width of the largest palm scale, to distal most lamella at tip of digits; lamellae under fourth digit of pes (LampIV). Morphometric data are given as % of SVL.
Morphometric analysis
We performed a multivariate analysis on morphometric variables collected from 19 specimens of the flaviventralis clade, which included eight specimens of C. ajijae and C. flaviventralis each and three specimens of the new species. We included only 15 of the 17 variables in this analysis because we were unable to obtain data for a few variables due to missing tails or digits (see Table
The phylogenetic analyses recovered largely similar topologies for the Maximum Likelihood (ML) and Bayesian analyses. However, several clades were unresolved in both analyses. The new species from Rajgad fort was sister to C. ajijae with strong support (Bootstrap: 96; Posterior probability: 1.0) and together formed a clade with C. flaviventralis with moderate support (Bootstrap: 79; Posterior probability: 0.9). This clade formed a strong sister relationship with other members of the girii clade (C. mahabali, C. amba, C. limayei, C. koynaensis and C. girii), thus we ascribe the new species, C. ajijae and C. flaviventralis to a separate flaviventralis clade. Within the girii clade, (C. girii + C. koynaensis) and C. limayei formed a strongly suppMaximum Likelihoodorted clade which together was sister to C. mahabali with strong support. Interestingly, our samples collected from Amba village were nested within C. mahabali.
Uncorrected pairwise sequence divergence indicated a deep genetic divergence in the 16S rRNA gene between the flaviventralis clade and the rest of the Cnemaspis (p-distance > 7.7%). Intraspecific genetic divergences estimated for members of the flaviventralis clade were ca. 0.7% for C. flaviventralis and 0–1.45% for C. ajijae (Table
Comparison of intraspecific and interspecific uncorrected p-distance for three closely related species of the Cnemaspis flaviventralis clade (C. flaviventralis, C. ajijae and C. rajgadensis sp. nov.) for the 16S rRNA gene. ‘N’ indicates number of pairwise comparisons.
Pair-wise comparisons | N | Mean (%) | Range (%) |
---|---|---|---|
Within C. flaviventralis | 1 | 0.72 | NA |
Within C. ajijae | 15 | 0.68 | 0–1.45 |
Within C. rajgadensis sp. nov. | 1 | 0.24 | NA |
C. flaviventralis – C. ajijae | 12 | 7.33 | 7.0–7.73 |
C. flaviventralis – C. rajgadensis sp. nov. | 4 | 7.25 | 7.0–7.49 |
C. ajijae – C. rajgadensis sp. nov. | 12 | 2.29 | 1.93–2.66 |
Principal component analysis (PCA) on the 15 variables indicates that the first three variables explained 85.9% of the variation (Table
Loadings obtained from the Principal Component Analysis on 15 morphometric variables collected for the Cnemaspis flaviventralis clade. Bold values indicate strong loading with correlation > 0.5.
Character | PC1 | PC2 | PC3 |
---|---|---|---|
SVL | 0.8825 | -0.0015 | -0.3526 |
TRL | 0.8656 | -0.0582 | -0.3186 |
TW | 0.6393 | -0.4388 | -0.5745 |
TLW | -0.0772 | 0.6070 | -0.7035 |
HL | 0.8347 | -0.2506 | 0.0132 |
HW | 0.8659 | 0.3021 | -0.1239 |
HD | 0.8458 | -0.0941 | -0.0452 |
FL | 0.8244 | 0.0044 | 0.1845 |
TBL | 0.9352 | -0.1007 | 0.1505 |
ED | 0.8005 | 0.4294 | 0.2876 |
EN | 0.9308 | 0.1877 | -0.0319 |
ES | 0.9418 | 0.2659 | 0.0089 |
ET | 0.8876 | 0.0467 | 0.3868 |
IN | 0.4838 | -0.7485 | -0.0733 |
IO | 0.8894 | 0.0548 | 0.2527 |
Eigen values | 9.8661 | 1.5932 | 1.4278 |
Standard deviation | 3.1410 | 1.2622 | 1.1949 |
Proportion of Variance | 0.6577 | 0.1062 | 0.0952 |
Cumulative Proportion | 0.6577 | 0.7640 | 0.8591 |
Family Gekkonidae Gray, 1825
Genus Cnemaspis Strauch, 1887
Holotype
: India • ♂, adult; Rajgad fort, Pune District, Maharashtra State; 18°14'46N, 73°40'55E; 1324 m a.s.l.; 27 Sept. 2020; Amit Sayyed leg.;
Paratypes
: India • 2 ♀, adults; same data as for holotype; Abhijit Nale, Kiran Ahire and Mahesh Bandgar leg.;
A small-sized Cnemaspis with adult SVL < 27 mm,; 7–7 supralabials; 6–7 infralabials; dorsal scales heterogeneous with small, granular, weakly keeled scales, intermixed with randomly arranged, weakly keeled, slightly larger tubercles; conical and spine-like tubercles absent on either side of the flanks but a row of enlarged tubercles present on the lower flanks; dorsal paravertebral scales 80–92; mid-dorsal scales 62–67; ventral surface of neck, pectoral, abdominal region, under limbs, and tail weakly keeled; mid-ventral scales 123–141; ventral scales across mid-body 28–29; subdigital lamellae under fourth digit of manus 15, under fourth digit of pes 18; males with 3 femoral pores and absence of precloacal pores; dorsal scales of tail granular, weakly keeled, roughly same in size and shape to those on mid-body dorsum, gradually becoming larger, flattened, blunt, sub-imbricate posteriorly, intermixed with whorls of slightly enlarged, strongly keeled tubercles; scales on ventral aspect of original tail imbricate, weakly keeled, without a series of enlarged sub-caudal scales, roughly same in size of those on dorsal tail; a single enlarged postcloacal spur on each side.
Cnemaspis rajgadensis sp. nov. differs from all other Indian species of Cnemaspis by having the following suite of characters: dorsal scales heterogeneous (vs. homogeneous in C. adii Srinivasulu et al., 2015, C. assamensis Das & Sengupta, 2000, C. boiei Gray, 1842, C. indica Gray, 1846, C. jerdonii Theobald, 1868, C. kolhapurensis Giri et al., 2009, C. littoralis Jerdon, 1853, C. mysoriensis Jerdon, 1853, C. nilagirica Manamendra et al., 2007, C. palakkadensis
C. rajgadensis sp. nov. closely resembles members of the girii and flaviventralis clade, all of which are small-sized species (SVL < 40 mm), have heterogeneous mid-dorsal scales, 2–5 femoral pores in males and having median subcaudals not enlarged. However, the new species can be differentiated from members of the girii clade by its higher number of ventral scales (28–29) across mid-body (vs. 22–24 in C. amba, 26–28 in C. girii, 20–26 in C. koynaensis, 26–27 in C. limayei and 26–27 in C. mahabali), absence of spine-like tubercles on flanks (vs. presence of spine-like tubercles in C. koynaensis), presence of enlarged tubercles row on lower flanks (vs. absence of tubercles on lower flanks of C. girii, C. limayei, C. mahabali), and by the weakly keeled ventral scales (vs. smooth in all members of the girii clade except C. mahabali).
The new species can be differentiated from C. ajijae and C. flaviventralis by its adult size (< 27 mm SVL vs. > 30 mm in C. ajijae and C. flaviventralis); short and wider than long head [HL 16.6% of SVL, HW 110.6% of HL] (vs. elongated in C. flaviventralis [HL22% of SVL, HW 81.8% of HL]); small eyes [ED 11.4% of HL] (vs. larger eyes in C. flaviventralis [ED 18.9% of HL] and C. ajijae [ED 21.6% of HL]); short snout [ES 68.7% of HL] (vs. long snout in C. ajijae [ES 82.4% of HL]); short and broad trunk [TW 61.5% of TRL](vs. slender in C. ajijae [TW 50.8% of TRL]). C. rajgadensis sp. nov. can also be differentiated by the weakly keeled ventral scales as opposed to the smooth scales in C. ajijae and C. flaviventralis.
Phylogentic relationships showing the topology from the Bayesian analyses for Indian species of Cnemaspis based on 16S rRNA gene. Values along the nodes are bootstrap support values from the Maximum Likelihood analysis and posterior probabilities from the Bayesian analyses respectively. The lineage in red indicates the phylogenetic cluster of the new species.
Adult male generally in good state of preservation, albeit slightly dehydrated (Fig.
Morphospace occupied by members of the flaviventralis clade including Cnemaspis rajgadensis sp. nov. as obtained from the Principal Component Analysis (PCA) along with distribution of the species. A Plot of PC1 and PC2, B Plot of PC1 and PC3, C – Map showing the type localities of C. rajgadensis sp. nov., C. ajijae and C. flaviventralis from northern Western Ghats of India.
(Fig.
(Fig.
The species epithet is derived from the Rajgad fort of Pune District, Maharashtra, India, from where the type series was collected. Rajgad is a hill fort and historical place, which was the capital of the Maratha Empire under the rule of Chhatrapati Shivaji Maharaj for almost 26 years during the 17th century. Previously known as Murumdev, Rajgad fort is located around 60 km to the south-west of Pune and about 15 km west of Nasrapur village in the range of northern Western Ghats.
Rajgad Dwarf Gecko
Currently, Cnemaspis rajgadensis sp. nov., is known from only its type locality – Rajgad fort, Pune District, Maharashtra.
The new species was found on the walls of old stone structure of Rajgad fort (Fig.
Cnemaspis rajgadensis sp. nov. holotype, adult male, SVL 25.7 mm (
Cnemaspis rajgadensis sp. nov. holotype, adult male, SVL 25.7 mm (
Habitat of Cnemaspis rajgadensis sp. nov. from the type locality at Rajgad fort, Pune, Maharashtra, India.
Mensural data for the type series of Cnemaspis rajgadensis sp. nov. *= gravid female, ^ = broken tail, – = pores not present. Measurements are in mm.
Measurements | Holotype ( |
Paratype ( |
Paratype ( |
---|---|---|---|
Sex | male | Female*^ | Female* |
SVL | 25.7 | 26.8 | 26.3 |
AG | 10.0 | 12.1 | 11.5 |
TW | 6.0 | 7.5 | 7.2 |
ED | 0.5 | 0.5 | 0.5 |
EN | 2.5 | 2.6 | 2.6 |
ES | 2.9 | 3.0 | 3.1 |
ET | 2.3 | 2.3 | 2.3 |
IN | 1.0 | 1.1 | 1.1 |
EOD | 0.2 | 0.2 | 0.2 |
HL | 4.3 | 4.3 | 4.5 |
HW | 4.7 | 4.8 | 5.0 |
HD | 2.9 | 3.3 | 3.6 |
IO | 2.0 | 2.3 | 3.2 |
UHL | 3.0 | 3.4 | 3.4 |
FAL | 4.2 | 3.9 | 3.5 |
PAL | 4.0 | 4.0 | 3.9 |
PAL | 4.0 | 4.0 | 3.9 |
FL1 | 1.6 | 1.6 | 1.4 |
FL2 | 1.9 | 2.0 | 2.0 |
FL3 | 2.2 | 2.3 | 2.3 |
FL4 | 2.4 | 3.0 | 2.9 |
FL5 | 2.1 | 2.2 | 2.2 |
FEL | 4.6 | 4.3 | 4.3 |
TBL | 3.4 | 4.0 | 3.9 |
TOL1 | 1.0 | 1.1 | 1.3 |
TOL2 | 2.3 | 2.0 | 1.9 |
TOL3 | 3.5 | 4.1 | 3.6 |
TOL4 | 4.3 | 4.6 | 3.9 |
TOL5 | 3.3 | 2.7 | 2.2 |
TL | 27.6 | 28.9 | 29.8 |
Morphometric and meristic data of the type series of Cnemaspis rajgadensis sp. nov. *= gravid female, ^ = broken tail, – = pores not present.
Character | Holotype ( |
Paratype ( |
Paratype ( |
---|---|---|---|
Sex | male | Female*^ | Female* |
SupL R/L | 7/7 | 7/7 | 7/7 |
InfL R/L | 7/7 | 6/6 | 7/7 |
SuS | 15 | 16 | 14 |
InO | 27 | 29 | 28 |
BeT | 18 | 17 | 17 |
PoN | 2 | 2 | 2 |
PoM | 2 | 2 | 2 |
PoP | 8 | 9 | 9 |
SuN | 3 | 3 | 3 |
CaS | 13 | 13 | 12 |
PvS | 80 | 92 | 86 |
MbS | 62 | 67 | 65 |
MvS | 123 | 127 | 141 |
BlS | 29 | 28 | 29 |
FPores | 3/3 | – | – |
PS | 25 | – | – |
MLam R | 9/12/15/15/12 | 10/12/15/15/14 | 9/12/15/15/13 |
PLam R | 8/13/18/18/18 | 9/13/18/18/18 | 9/13/16/18/18 |
The topology of our phylogenetic tree based on the 16S rRNA gene was more similar to that of
The new species, Cnemaspis rajgadensis sp. nov., would be the twelfth species of the genus Cnemaspis described from the northern Western Ghats, India, thus underlining the endemic diversity of this region. This study also marks the importance of forts in Maharashtra as potential habitats for micro fauna. Although these forts are man-made intrusions into natural habitats, these sites being of archaeological importance, are currently protected by law from extensive infrastructure and developmental activities. Most of these forts being situated in the rugged terrains of the northern Western Ghats remain inaccessible to extensive human activities and potentially have conservation implications in protecting local flora and fauna. The current study, as well as the recently described C. ranganaensis from Rangana fort, near Kolhapur, Maharashtra indicate rich bio-diversity of endemic reptile fauna occupying these forts. These forts have always been protected for their archaeological importance, however have never been considered for conservation due to lack of data. Dedicated biodiversity surveys of these forts in Maharashtra is much needed to highlight the importance of these structures and also draft appropriate conservation plans.
We thank all the reviewers for their constructive comments, which greatly improved the manuscript. We thank the Forest Departments of Maharashtra for issuing collecting permits and for their support during field surveys. We also thank the Institute of Natural History Education and Research (INHER), Pune for institutional facilities and partial funding received for this study. We thank Gaurang Gowande and Shubhankar Deshpande for their help in this study. We are grateful to President, INHER, Pune for the encouragement. We thank Deepak Apte and Rahul Khot, of the Bombay Natural History Society, Mumbai for providing access to specimens and registration of the type specimens. We also thank Vithoba Hegde