Abstract
In this study, we re-examine the Darevskia parvula group comprehensively using morphology, osteology and mitochondrial phylogeny, and describe a new endemic species from Turkey: Darevskia tuniyevi sp. nov. A total of 257 adult specimens were evaluated for external morphology (scalation and biometry) with univariate (descriptive statistics and ANOVA with post-hoc tests) and multivariate (Discriminant Analysis and ANOSIM) analyses. In parallel, osteological data and molecular analyses using three DNA markers (mitochondrial 16S rRNA and Cyt-b, nuclear Rag-1) were used to complete the description of the new taxon. The molecular phylogenetic analyses indicated that the D. parvula group is composed of three taxa as D. parvula, D. adjarica and D. tuniyevi sp. nov., and showed that D. adjarica and D. tuniyevi sp. nov. are reciprocal sister taxa. On the other hand, D. adjarica is morphologically very different from other two forms, while D. parvula is hardly distinguishable externally from D. tuniyevi sp. nov. Therefore, we can consider that D. parvula and D. tuniyevi sp. nov. are cryptic species. These two cryptic species retain their primitive morphology within the group, while D. adjarica has changed, perhaps due to different bioclimatic conditions in its Pleistocene refuge and current area.
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Data availability
The molecular genetic datasets generated and/or analysed in the present study are available from the GenBank repository (https://www.ncbi.nlm.nih.gov/genbank/).
References
Ahmadzadeh, F., Flecks, M., Carretero, M. A., Mozaffari, O., Böhme, W., Harris, D. J., et al. (2013). Cryptic speciation patterns in Iranian Rock Lizards uncovered by integrative taxonomy. PLoS one, 8(12), e80563–e80617. https://doi.org/10.1371/journal.pone.0080563.
Arnold, E. N. (1973). Relationships of the Palaearctic lizards assigned to the genera Lacerta, Algyroides and Psammodromus (Reptilia: Lacertidae). Bulletin of the British Museum (Natural History) Zoology, London, 25(8), 289–366.
Arnold, E. N., Arribas, O. J., & Carranza, S. (2007). Systematics of the Palaearctic and Oriental lizard tribe Lacertini (Squamata: Lacertidae: Lacertinae), with descriptions of eight new genera. Zootaxa, 1430, 1–86. https://doi.org/10.11646/zootaxa.1430.1.1
Arribas, O., Ilgaz, Ç., & Kumlutaş, Y. (2018). Reevaluation of the intraspecific variability in Darevskia parvula (Lantz and Cyrén, 1913): an integrated approach using morphology, osteology and genetics (Squamata: Lacertidae). Zootaxa, 4472, 71–099. https://doi.org/10.11646/zootaxa.4472.1.3
Arribas, O. J. (2010). Intraspecific variability of the Carpetane Lizard (Iberolacerta cyreni [Müller & Hellmich, 1937]) (Squamata: Lacertidae), with special reference to the unstudied peripheral populations from the Sierras de Avila (Paramera, Serrota and Villafranca). Bonn Zoological Bulletin, 57(2), 197–210.
Blackiht, R. E., & Reyment, R. A. (1971). Multivariate morphometrics (p. 412). Academic Press.
Bodenheimer, F. S. (1944). Introduction into the knowledge of the Amphibia and Reptilia of Turkey. Revue de la Faculté des Sciences de l’Universite d’Istanbul. Ser. b., 9, 1–78.
Candan, K., Kornilios, P., Ayaz, D., Kumlutaş, Y., Gül, S., Yıldırım-Caynak, E., & Ilgaz, Ç. (2021). Cryptic genetic structure within Valentin’s Lizard, Darevskia valentini (Boettger, 1892) (Squamata, Lacertidae), with implications for systematics and origins of parthenogenesis. Systematics and Biodiversity. https://doi.org/10.1080/14772000.2021.1909171
Carranza, S., & Arnold, E. N. (2012). A review of the geckos of the genus Hemidactylus (Squamata: Gekkonidae) from Oman based on morphology, mitochondrial and nuclear data, with descriptions of eight new species. Zootaxa, 3378, 1–95. https://doi.org/10.11646/zootaxa.3378.1.1
Carstens, B. C., Pelletier, T. A., Reid, N. M., & Satler, J. D. (2013). How to fail at species delimitation. Molecular Ecology, 22(17), 4369–4383. https://doi.org/10.1111/mec.12413
Chernomor, O., von Haeseler, A., & Minh, B. Q. (2016). Terrace aware data structure for phylogenomic inference from supermatrices. Systematic Biology, 65(6), 997–1008. https://doi.org/10.1093/sysbio/syw037
Clark, R. J., & Clark, E. D. (1973). Collection of Amphibians and Reptiles from Turkey. Occasional Papers of the California Academy of Sciences, 104, 1–62.
Clarke, K. R. (1988). Detecting change in benthic community structure.In: Oger, R (ed), Proceedings of invited papers, 14th international biometric conference, Namour, Belgium, 131–142.
Clarke, K. R. (1993). Non-parametric multivariate analyses of changes in community structure. Australian Journal of Ecology, 18, 117–143. https://doi.org/10.1111/j.1442-9993.1993.tb00438.x
Clement, M., Posada, D., & Crandall, K. A. (2000). TCS: A computer program to estimate gene genealogies. Molecular Ecology, 9(10), 1657–1660. https://doi.org/10.1046/j.1365-294x.2000.01020.x
Darevsky, I. S. (1967). Rock lizards of the Caucasus (Systematics, Ecology and Phylogenesis of the polymorphic groups of Rock lizards of the Subgenus Archaeolacerta). Nauka press. Leningrad. 216 pp [Translation: New Delhi: Indian National Scientific Documentation Centre, 276 pp].
Darevsky, I. S., & Eiselt, J. (1980). Neue Felseneidechsen (Reptilia: Lacertidae) aus dem Kaukasus und aus der Türkei. Amphibia-Reptilia, 1(1), 29–40.
Felsenstein, J. (1985). Confidence limits on phylogenies: An approach using the bootstrap. Evolution; International Journal of Organic Evolution, 39(4), 783–791. https://doi.org/10.1111/j.1558-5646.1985.tb00420.x
Freitas, S., Rocha, S., Campos, J., Ahmadzadeh, F., Corti, C., Sillero, N., Ilgaz, Ç., Kumlutaş, Y., Arakelyan, M., Harris, D. J., & Carretero, M. A. (2016a). Parthenogenesis through the ice ages: A biogeographic analysis of Caucasian rock lizards (genus Darevskia). Molecular Phylogenetics and Evolution, 102, 117–127. https://doi.org/10.1016/j.ympev.2016.05.035
Freitas, S., Vavakou, A., Arakelyan, M., Drovetski, S. V., Crnobrnja-Isailovic, J., Kidov, A. A., Cogalniceanu, D., Corti, C., Lymberakis, P., Harris, D. J., & Carretero, M. A. (2016b). Cryptic diversity and unexpected evolutionary patterns in the meadow lizard, Darevskia praticola (Eversmann, 1834). Systematics and Biodiversity, 14(2), 184–197. https://doi.org/10.1080/14772000.2015.1111267
Guindon, S., Dufayard, J. F., Lefort, V., Anisimova, M., Hordijk, W., & Gascuel, O. (2010). New algorithms and methods to estimate maximum-likelihood phylogenies: Assessing the performance of PhyML 3.0. Systematic Biology, 59(3), 307–321. https://doi.org/10.1093/sysbio/syq010
Gündüz, I., Jaarola, M., Tez, C., Yeniyurt, C., Polly, P. D., & Searle, J. B. (2007). Multigenic and morphometric differentiation of ground squirrels (Spermophilus, Sciuridae, Rodentia) in Turkey, with a description of a new species. Molecular Phylogenetics and Evolution, 43, 916–935. https://doi.org/10.1016/j.ympev.2007.02.021
Gür, H. (2016). The Anatolian diagonal revisited: Testing the ecological basis of a biogeographic boundary. Zoology in the Middle East, 62(3), 189–199. https://doi.org/10.1080/09397140.2016.1226544
Hall, T. A. (1999). BioEdit: A User-Friendly Biological Sequence Alignment Editor and Analysis Program for Windows 95/98/NT. Nucleic Acids Symposium Series, 41, 95–98.
Hintze, J. (2007). NCSS, PASS and GESS.Number Cruncher Statistical Systems. Kaysville, Utah. (Available from www.NCSS.com)
Ilgaz, Ç. (2009). Comparative morphology of Darevskia parvula (Lantz-Cyren 1936) (Sauria: Lacertidae) subspecies in Northeastern Anatolia. Turkey. North-West Journal of Zoology, 5(2), 263–280.
Jablonski, D., Kukushkin, O. V., Avcı, A., Bunyatova, S., Kumlutaş, Y., Ilgaz, Ç., Polyakova, E., Shiryaev, K., Tuniyev, B., & Jandzik, D. (2019). The biogeography of Elaphe sauromates (Pallas, 1814), with a description of a new rat snake species. PeerJ, 7, e6944. https://doi.org/10.7717/peerj.6944
Kalyaanamoorthy, S., Minh, B. Q., Wong, T. K. F., von Haeseler, A., & Jermiin, L. S. (2017). ModelFinder: Fast model selection for accurate phylogenetic estimates. Nature Methods, 14(6), 587–589. https://doi.org/10.1038/nmeth.4285
Kapli, P., Botoni, D., Ilgaz, Ç., Kumlutaş, Y., Avcı, A., Rastegar-Pouyani, N., Fathinia, B., Lymberakis, P., Ahmadzadeh, F., & Poulakakis, N. (2013). Molecular phylogeny and historical biogeography of the Anatolian lizard Apathya (Squamata, Lacertidae). Molecular Phylogenetics and Evolution, 66(3), 992–1001. https://doi.org/10.1016/j.ympev.2012.12.002
Kapli, P., Lutteropp, S., Zhang, J., Kobert, K., Pavlidis, P., Stamatakis, A., & Flouri, T. (2017). Multi-rate Poisson tree processes for singlelocus species delimitation under maximum likelihood and Markov chain Monte Carlo. Bioinformatics, 33(11), 1630–1638. https://doi.org/10.1093/bioinformatics/btx025
Karakasi, D., Ilgaz, Ç., Kumlutaş, Y., Candan, K., Güçlü, Ö., Kankılıç, T., Beşer, N., Sindaco, R., Lymberakis, P., & Poulakakis, N. (2021). More evidence of cryptic diversity in Anatololacerta species complex Arnold, Arribas and Carranza, 2007 (Squamata: Lacertidae) and re-evaluation of its current taxonomy. Amphibia-Reptilia, 42(2), 201–216. https://doi.org/10.1163/15685381-bja10045
Katoh, K., & Standley, D. M. (2013). MAFFT multiple sequence alignment software version 7: Improvements in performance and usability, outlines version 7. Molecular Biology and Evolution, 30(4), 772–780. https://doi.org/10.1093/molbev/mst010
Köhler, G. (2012). Color Catalogue for Field Biologists. Herpeton ed., Offenbach. 49 pp.
Korkmaz, E. M., Lunt, D. H., Çıplak, B., Değerli, N., & Başıbüyük, H. H. (2014). The contribution of Anatolia to European phylogeography: The centre of origin of the meadow grasshopper, Chorthippus parallelus. Journal of Biogeography, 41, 1793–1805. https://doi.org/10.1111/JBI.12332
Kornilios, P., Jablonski, D., Sadek, R. A., Kumlutaş, Y., Olgun, K., Avci, A., & Ilgaz, Ç. (2020). Multilocus species-delimitation in the Xerotyphlops vermicularis (Reptilia: Typhlopidae) species complex. Molecular Phylogenetics and Evolution, 152, 106922. https://doi.org/10.1016/j.ympev.2020.106922
Kornilios, P., Kumlutaş, Y., Lymberakis, P., & Ilgaz, Ç. (2018). Cryptic diversity and molecular systematics of the Aegean Ophiomorus skinks (Reptilia: Squamata), with the description of a new species. Journal of Zoological Systematics and Evolutionary Research, 56(3), 364–381. https://doi.org/10.1111/jzs.12205
Kumar, S., Stecher, G., Li, M., Knyaz, C., Tamura, K. (2018). MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms. Molecular Biology and Evolution, 35, 1547–1549. https://doi.org/10.1093/molbev/msy096
Kurnaz, M., Kutrup, B., Hosseinian Yousefkhani, S. S., Koc, H., Bülbül, U., & Eroğlu, A. İ. (2019). Phylogeography of the red-bellied lizard, Darevskia parvula in Turkey. Mitochondrial DNA Part A, 30(3), 556–566. https://doi.org/10.1080/24701394.2019.1580270
Lantz, L. A., & Cyrén, O. (1913). Eine neue Varietät der Felseneidechse Lacerta saxicola Eversmann parvula nov. var. Mitteilungen Kaukasus Museum, 7(2), 163–168.
Legendre, P., & Legendre, L. (1998). Numerical Ecology (p. 853). Elsevier Science B. V.
Leigh, J. W., & Bryant, D. (2015). PopART: Full-feature software for haplotype network construction. Methods in Ecology and Evolution, 6(9), 1110–1116. https://doi.org/10.1111/2041-210X.12410
Lemey, P., Salemi, M., & Vandamme, A. N. (2009). The Phylogenetic Handbook (Second edition).
Manvelidze, Z., Eminagaoglu, O., Memiadze, N. V., & Charazishvili, DSh. (2009). Diversity of endemic plant species of adjara-shavsat florist region. Annals of Agrarian Science., 7, 150–152.
Minh, B. Q., Nguyen, M. A., & von Haeseler, A. (2013). Ultrafast approximation for phylogenetic bootstrap. Molecular Biology and Evolution, 30(5), 1188–1195. https://doi.org/10.1093/molbev/mst024
Mittermeier, R. A., Gil, P. R., Hoffmann, M., Pilgrim, J., Brooks, T., Mittermeier, C. G., Lamoreux, J., & Fonseca, G. A. B. (2004). Hotspots revisited: Earth’s biologically richest and most endangered terrestrial ecoregions. CEMEX.
Murtskhvaladze, M., Tarkhnishvili, D., Anderson, C. L., & Kotorashvili, A. (2020). Phylogeny of caucasian rock lizards (Darevskia) and other true lizards based on mitogenome analysis: Optimisation of the algorithms and gene selection. PLoS one, 15(6).
Mutun, S. (2010). Intraspecific genetic variation and phylogeography of the oak gallwasp Andricus caputmedusae (Hymenoptera: Cynipidae): Effects of the Anatolian diagonal. Acta Zoologica Academiae Scientiarum Hungaricae, 56, 153–172.
Psonis, N., Antoniou, A., Kukushkin, O., Jablonski, D., Petrov, B., Crnobrnja-Isailović, J., Sotiropoulos, K., Gherghel, I., Lymberakis, P., & Poulakakis, N. (2017). Hidden diversity in the Podarcis tauricus (Sauria, Lacertidae) species subgroup in the light of multilocus phylogeny and species delimitation. Molecular Phylogenetics and Evolution, 106, 6–17. https://doi.org/10.1016/j.ympev.2016.09.007
Rambaut, A., Drummond, A. J., Xie, D., Baele, G., & Suchard, M. A. (2018). Posterior summarisation in Bayesian phylogenetics using Tracer 1.7. Systematic Biology, 67(5), 901–904. https://doi.org/10.1093/sysbio/syy032
Rawat, U. S., & Agarwall, N. K. (2015). Biodiversity: Concept, threats and conservation. Environment Conservation Journal, 16(3), 19–28.
Rokas, A., Atkinson, R. J., Webster, L., Csóka, G., & Stone, G. N. (2003). Out of Anatolia: Longitudinal gradients in genetic diversity support an eastern origin for a circum-Mediterranean oak gallwasp Andricus quercustozae. Molecular Ecology, 12, 2153–2174. https://doi.org/10.1046/j.1365-294X.2003.01894.x
Ronquist, F., Teslenko, M., Van Der Mark, P., Ayres, D. I., Darling, A., Hohna, S., Larget, B., Liu, L., Suchard, M. A., & Huelsenbeck, J. P. (2012). MrBayes 3.2: Efficient Bayesian phylogenetic inference and model choice across alarge model space. Systematic Biology, 61(3), 539–542. https://doi.org/10.1093/sysbio/sys029
Rozas, J., Ferrer-Mata, A., Sanchez-DelBarrio, J. C., Guirao-Rico, S., Librado, P., Ramos-Onsins, S. E., & Sanchez-Gracia, A. (2017). DnaSP 6: DNA sequence polymorphism analysis of large data sets. Molecular Biology and Evolution, 34(12), 3299–3302. https://doi.org/10.1093/molbev/msx248
Seaby, R. M. H., & Henderson, P. A. (2019). Community Analysis Package 6.0. Pisces Conservation Ltd, Lymington, UK. 164 pp. (Available from www.pisces-conservation.com)
Şekercioğlu, Ç. H., Anderson, S., Akçay, E., Bilgin, R., Can, Ö. E., Semiz, G., Tavşanoğlu, Ç., Yokeş, M. B., Soyumert, A., Sağlam, İK., Yücel, M., & Dalfes, H. N. (2011). Turkey’s globally important biodiversity in crisis. Biological Conservation, 144, 2752–2769. https://doi.org/10.1016/j.biocon.2011.06.025
Simó-Riudalbas, M., Metallinou, M., de Pous, P., Els, J., Jayasinghe, S., Péntek-Zakar, E., et al. (2017). Cryptic diversity in Ptyodactylus (Reptilia: Gekkonidae) from the northern Hajar Mountains of Oman and the United Arab Emirates uncovered by an integrative taxonomic approach. PLoS one, 12.
Sindaco, R., Kornilios, P., Sacchi, R., & Lymberakis, P. (2014). Taxonomic reassessment of Blanus strauchi (Bedriaga, 1884) (Squamata: Amphisbaenia: Blanidae), with the description of a new species from southeast Anatolia (Turkey). Zootaxa, 3795(3), 311–326. https://doi.org/10.11646/zootaxa.3795.3.6
Sokal, R. R., & Rohlf, J. (1969). Biometry. The principles and practice of statistics in Biological research. W.F. Freeman and C., New York, 776 pp.
Stephens, M., & Scheet, P. (2005). Accounting for decay of linkage disequilibrium in haplotype inference and missingdata imputation. American Journal of Human Genetics, 76(3), 449–462. https://doi.org/10.1086/428594
Stephens, M., Smith, N. J., & Donnelly, P. (2001). A new statistical method for haplotype reconstruction from population data. American Journal of Human Genetics, 68(4), 978–989. https://doi.org/10.1086/319501
Suchard, M. A., Lemey, P., Baele, G., Ayres, D. L., Drummond, A. J., & Rambaut, A. (2018). Bayesian phylogenetic and phylodynamic data integration using BEAST 1.10. Virus Evolution, 4(1), vey016. https://doi.org/10.1093/ve/vey016
Tamar, K., Carranza, S., In den Bosch, H., Sindaco, R., Moravec, J., & Meiri, S. (2015). Hidden relationships and genetic diversity: Molecular phylogeny and phylogeography of the Levantine lizards of the genus Phoenicolacerta (Squamata: Lacertidae). Molecular Phylogenetics and Evolution, 91, 86–97. https://doi.org/10.1016/j.ympev.2015.05.002
Tamar, K., Mitsi, P., & Carranza, S. (2019). Cryptic diversity revealed in the leaf-toed gecko Asaccus montanus (Phyllodactylidae) from the Hajar Mountains of Arabia. Journal of Zoological Systematics and Evolutionary Research, 57(2), 369–382. https://doi.org/10.1111/jzs.12258
Tarkhnishvili, D., Murtskhvaladze, M., & Gavashelishvili, A. (2013). Speciation in Caucasian lizards: Climatic dissimilarity of the habitats is more important than isolation time. Biological Journal of the Linnean Society, 109(4), 876–892. https://doi.org/10.1111/bij.12092
Tarkhnishvili, D., Yanchukov, A., Şahin, M. K., Gabelaia, M., Murtskhvaladze, M., Candan, K., Galoyan, E., Arakelyan, M., Iankoshvili, G., Kumlutaş, Y., Ilgaz, Ç., Matur, F., Çolak, F., Erdolu, M., Kurdadze, S., Barateli, N., & Anderson, C. (2020). Genotypic similarities among the parthenogenetic rock lizards Darevskia with presumed different hybrid origins. BMC Evolutionary Biology, 20, 122. https://doi.org/10.1186/s12862-020-01690-9
Tavşanoğlu, Ç. (2016). Anadolu’nun yüksek biyoçeşitliliği: evrim bunun neresinde? Akış, I. ve Altınışık, N.E. (Editörler) Yazılama Yayınevi, İstanbul, s. 207–225.
Trifinopoulos, J., Nguyen, L. T., Von Haeseler, A., & Minh, B. Q. (2016). W-IQ-TREE: A fast online phylogenetic tool for maximum likelihood analysis. Nucleic Acids Research, 44(W1), 232–235. https://doi.org/10.1093/nar/gkw256
Vamberger, M., Stuckas, H., Ayaz, D., Gracia, E., Aloufi, A. A., Els, J., Mazanaeva, L. F., Kami, H. G., & Fritz, U. (2013). Conservation genetics and phylogeography of the poorly known Middle Eastern terrapin Mauremys caspica (Testudines: Geoemydidae). Organisms Diversity and Evolution, 13, 77–85. https://doi.org/10.1007/s13127-012-0102-6
Zhang, J., Kapli, P., Pavlidis, P., & Stamatakis, A. (2013). A General Species Delimitation Method with Applications to Phylogenetic Placements. Bioinformatics, 29(22), 2869–2876. https://doi.org/10.1093/bioinformatics/btt499
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We would like to thank Dr. David Tarkhnishvili for valuable comments on an earlier version of the manuscript and for providing us with Darevskia adjarica samples from Georgia. We also thank Catherine YiÄŸit for linguistic help with the text, and the reviewers for their many precious comments and suggestions to improve the manuscript.
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Arribas, O., Candan, K., Kurnaz, M. et al. A new cryptic species of the Darevskia parvula group from NE Anatolia (Squamata, Lacertidae). Org Divers Evol 22, 475–490 (2022). https://doi.org/10.1007/s13127-022-00540-4
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DOI: https://doi.org/10.1007/s13127-022-00540-4