hybridization; multilocus barcoding; phylogeography; reptiles; taxonomy; Ecology, Evolution, Behavior and Systematics; Animal Science and Zoology
Abstract :
[en] With five currently recognized species that form several secondary contact zones, slow worms (Anguidae: Anguis) offer a valuable model to study the fate of evolutionary lineages in the face of hybridization and genetic introgression. The relationships between the Western Slow Worm Anguis fragilis and the Italian Slow Worm Anguis veronensis are particularly puzzling. Their respective distributions remain poorly known on the edges of their parapatric ranges, as both species lack external differentiation. Contra earlier mitochondrial phylogenies, new phylogenomic inferences have shown that A. fragilis and A. veronensis are sister taxa, thus casting doubts on their specific status. In this study, we analyze the A. fragilis/veronensis transition in southeastern France, based on one mitochondrial (ND2) and two nuclear (PRLR and HA1) genetic markers in 81 specimens from 61 localities. The ranges of A. fragilis and A. veronensis roughly extends northwest and southeast of the Rhône-Durance valleys, respectively, with clear signs of introgressive hybridization in the areas of contact (notably the eastern parts of the lower Rhône valley). Based on the three molecular markers analyzed, gene flow does not seem to reach outside the narrow hybrid zone, which likely indicates (incomplete) intrinsic reproductive isolation. Hence, we provisionally suggest maintaining A. veronensis as a separate species from A. fragilis. More generally, patterns of genetic divergence, external differentiation, and hybridization (both historical and contemporary) in Anguis ssp. supports a speciation continuum spanning from cryptic, genetically compatible alloparapatric lineages to phenotypically distinct, deeply diverged and fully reproductively isolated taxa able to coexist in sympatry.
Dufresnes, Christophe ; Laboratory for Amphibian Systematic & Evolutionary Research, College of Biology & the Environment, Nanjing Forestry University, Nanjing, China
Sourrouille, Patricia; CEFE, CNRS, Univ. Montpellier, EPHE, IRD, Montpellier, France
Olivier, Anthony; Tour du Valat, Research Institute for the Conservation of Mediterranean Wetlands, Arles, France
Ballouard, Jean-Marie; CRCC, Centre for Research and Conservation of Chelonians, SOPTOM, Var, Carnoules, France
Leccia, Marie-France; Parc National du Mercantour, Nice, France
Tiné, Rémi; Syndicat Mixte Camargue Gardoise, Centre du Scamandre, Route des Iscles, Gallician, Vauvert, France
Cheylan, Marc; Biogéographie et Ecologie des Vertébrés, CEFE, EPHE-PSL University, Univ. Montpellier, CNRS, IRD, Montpellier, France
Le Henanff, Maxime; ECO-MED - Ecologie & Médiation, Résidence Atrium, Montpellier, France
Nicolas, Jean; Camplong, France
Caron, Sébastien; CRCC, Centre for Research and Conservation of Chelonians, SOPTOM, Var, Carnoules, France
Massez, Grégoire; Les Amis des Marais du Vigueirat, Les Marais du Vigueirat, France
Levasseur, Romain; CRCC, Centre for Research and Conservation of Chelonians, SOPTOM, Var, Carnoules, France ; Le Val, France
Pille, Fabien ; Université de Liège - ULiège > Freshwater and OCeanic science Unit of reSearch (FOCUS) ; CRCC, Centre for Research and Conservation of Chelonians, SOPTOM, Var, Carnoules, France
Peyre, Olivier; PANDORA, Avignon Cédex 9, France
Thibault, Marc; Tour du Valat, Research Institute for the Conservation of Mediterranean Wetlands, Arles, France
Crottini, Angelica; CIBIO/InBio, Centro de Investigação em Biodiversidade e Recursos Genéticos Campus Agrário de Vairão, Vairão, Portugal ; Departamento de Biologia, Faculdade de Ciências, Universidade do Porto, Porto, Portugal ; BIOPOLIS Program in Genomics, Biodiversity and Land Planning, CIBIO, Vairão, Portugal
Fuento, Nicolas; LPO PACA, Hyères, France
Crochet, Pierre-André; CEFE, CNRS, Univ. Montpellier, EPHE, IRD, Montpellier, France
Exploring the speciation continuum of slow worms: location and extent of the Anguis fragilis/veronensis hybrid zone in southeastern France
Publication date :
2023
Journal title :
Amphibia-Reptilia
ISSN :
0173-5373
eISSN :
1568-5381
Publisher :
Brill Academic Publishers
Peer reviewed :
Peer Reviewed verified by ORBi
Funding text :
We are grateful to F. Arnaboldi, D. Arsovski, J.-M. Ballouard / SOPTOM, H. Brosius, F. Chapelur, S. Combeaud, P. Coursier-Fournial, V. Danias, B. Delahaie, F. Dhermain, E. Didner, J. Dublon, G. Gillet, C. Jakob / Ecole domaine du possible, T. Lebard, I. Lhommedet, A. Lucas, J.-L. Lucchesi, A. Lyet, L. Malthieux, P. Migaud, A. Miralles, P. Ormea, B. Schatz, T. Schwartz, A. Rocha, L. Tillion, F. Veyrunes, M. Voronkoff, A. Westerström, L. Westrin and L. Zimmerman for their help with samples collection and to P. Geniez for his help with the samples curation. CD was supported by the Research Found for International Scientists (RFIS) of the National Science Foundation of China (NSFC) (N°3211101356), as well as the Taxon-Omics Priority Programme (SPP1991) of the Deutsche Forschungsgemeinschaft (DFG) (grant number VE247/19-1). AC was supported by the Portuguese National Funds through FCT– Fundação para a Ciência e a Tecnologia (research contract 2020.00823.CEECIND).
Arnaud-Fassetta, G. (2003): River channel changes in the Rhone Delta (France) since the end of the Little Ice Age: geomorphological adjustment to hydroclimatic change and natural resource management. Catena 51: 141-172.
Arntzen, J.W., de Vries, W., Canestrelli, D., Martínez-Solano, I. (2017): Hybrid zone formation and contrasting outcomes of secondary contact over transects in common toads. Molecular Ecology 26: 5663-5675.
Arntzen, J.W., Canestrelli, D., Martínez-Solano, I. (2020): Environmental correlates of the European common toad hybrid zone. Contribution to Zoology 89: 270-281.
Asztalos, M., Schultze, N., Ihlow, F., Geniez, P., Berroneau, M., Delmas, C., Guiller, G., Legentilhomme, J., Kindler, C., Fritz, U. (2020): How often do they do it? An in-depth analysis of the hybrid zone of two grass snake species (Natrix astreptophora and Natrix helvetica). Biological Journal of the Linnean Society 131: 756-773.
Barton, N.H., Hewitt, G.M. (1985): Analysis of hybrid zones. Annual Review of Ecology & Systematics 16: 113-148.
Beddek, M., Zenboudji-Beddek, S., Géniez, P., Fathalla, R., Sourouille, P., Arnal, V., Dellaoui, B., Koudache, F., Telailia, S., Peyre, O., Crochet, P.A. (2018): Comparative phylogeography of amphibians and reptiles in Algeria suggest common causes for the east-west phylogeographic breaks in the Maghreb. PLoS ONE 13: e0201218.
Benkovský, N., Moravec, J., Gvoždíková Javůrková, V.G., Šifrová, H., Gvoždík, V., Jandzik, D. (2021): Phenotypic differentiation of the slow worm lizards (Squamata: Anguis) across their contact zone in Central Europe. PeerJ 9: e12482.
Bernardo, P.H., Sánchez-Ramírez, S., Sánchez-Pacheco, S.J., Álvarez-Castañeda, S.T., Aguilera-Miller, E.F., Mendez-de la Cruz, F.R., Murphy, R.W. (2019): Extreme mito-nuclear discordance in a Peninsular lizard: the role of drift, selection, and climate. Heredity 123: 359-370.
Bravard, J.-P., Gaydou, P. (2015): Historical development and integrated management of the Rhône River floodplain, from the Alps to the Camargue delta, France. In: Geomorphic Approaches to Integrated Floodplain, pp. 289-320. Hudson, P.F., Middelkoop, H., Eds, Springer, New York.
Bruneton, H., Arnaud-Fassetta, G., Provansal, M., Sistach, D. (2001): Geomorphological evidence for fluvial change during the Roman period in the lower Rhone valley (southern France). Catena 45: 287-312.
Chenuil, A., Cahill, A.E., Délémontey, N., Du Salliant, E., Fanton, H. (2019): Problems and questions posed by cryptic species. A framework to guide future studies. In: From Assessing to Conserving Biodiversity. History, Philosophy and Theory of the Life Sciences, vol. 24, pp. 77-106. Casetta, E., Marques da Silva, J., Vecchi, D., Eds, Springer, Berlin, Germany.
Dufour, P., Pons, J.-M., Collinson, M., Hamza, A., Gernigon, J., Dies, J.I., Sourrouille, P., Crochet, P.-A. (2017): Multilocus barcoding confirms the occurrence of Elegant Terns in Western Europe. Journal of Ornithology 158: 351-361.
Dufresnes, C., Jablonski, S.N. (2022): A genomic revolution in amphibian taxonomy. Science 377: 6612.
Dufresnes, C., Litvinchuk, S.N. (2022): Diversity, distribution and molecular species delimitation in frogs and toads from the Eastern Palearctic. Zoological Journal of the Linnean Society 195: 695-760.
Dufresnes, C., Nicieza, A.G., Litvinchuk, S.N., Rodrigues, N., Jeffries, D.L., Vences, M., Perrin, N., Martínez-Solano, I. (2020a): Are glacial refugia hotspots of speciation and cyto-nuclear discordances? Answers from the genomic phylogeography of Spanish common frogs. Molecular Ecology 29: 986-1000.
Dufresnes, C., Pribille, M., Alard, B., Gonçalves, H., Amat, F., Crochet, P.-A., Dubey, S., Perrin, N., Fumagalli, L., Vences, M., Martínez-Solano, I. (2020b): Integrating hybrid zone analyses in species delimitation: lessons from two anuran radiations of the Western Mediterranean. Heredity 124: 423-438.
Dufresnes, C., Litvinchuk, S.N., Rozenblut-Kościsty, B., Rodrigues, N., Perrin, N., Crochet, P.-A., Jeffries, D.L. (2020c): Hybridization and introgression between toads with different sex chromosome systems. Evolution Letters 4: 444-456.
Dufresnes, C., Berroneau, M., Dubey, S., Litvinchuk, S.N., Perrin, N. (2020d): The effect of phylogeography history on species boundaries: a comparative framework in Hyla tree frogs. Scientific Reports 10: 5502.
Dufresnes, C., Brelsford, A., Jeffries, D.L., Mazepa, G., Suchan, T., Canestrelli, D., Nicieza, A., Fumagalli, L., Dubey, S., Martínez-Solano, I., Litvinchuk, S.N., Vences, M., Perrin, N., Crochet, P.-A. (2021): Mass of genes rather than master genes underlie the genomic architecture of amphibian speciation. Proceedings of the National Academy of Sciences USA 118: e2103963118.
Garnier, G., Fournial, P., Bonnet, X., Maxime, L., Caron, S., Ballouard, J.-M. (2021): Première observation d'Anguis sp. sur l'île de Porquerolles (Var, France). Bulletin de la société herpétologique de France 178: 80-81.
Gvoždík, V., Jandzik, D., Lymberakis, P., Jablonski, D., Moravec, J. (2010): Slow worm, Anguis fragilis (Reptilia: Anguidae) as a species complex: genetic structure reveals deep divergences. Molecular Phylogenetics & Evolution 55: 460-472.
Gvoždík, V., Benkovsky, N., Crottini, A., Bellati, A., Moravec, J., Romano, A., Sacchi, R., Jandzik, D. (2013): An ancient lineage of slow worms, genus Anguis (Squamata: Anguidae), survived in the Italian Peninsula. Molecular Phylogenetics & Evolution 69: 1077-1092.
Gvoždík, V., Nečas, T., Jablonski, D., Moriarty Lemmon, E., Lemmon, A.R., Jandzik, D., Moravec, J. (2023): Phylogenomics of Anguis and Pseudopus (Squamata, Anguidae) indicates Balkan-Apennine mitochondrial capture associated with the Messinian event. Molecular Phylogenetics & Evolution 180: 107674.
Hebert, P.D.N., Cywinska, A., Ball, S.L., Jeremy, R. (2003): Biological identifications through DNA barcodes. Proceedings of the Royal Society B: Biological Sciences. 270: 313-321.
Hewitt, G.M. (1988): Hybrid zones - natural laboratories for evolutionary studies. Trends in Ecology & Evolution 3: 158-167.
Hinojosa, J.C., Koubínová, D., Szenteczki, M.A., Pitteloud, C., Dincă, V., Alvarez, N., Vila, R. (2019): A mirage of cryptic species: genomics uncover striking mitonuclear discordance in the butterfly Thymelicus sylvestris. Molecular Ecology 28: 3857-3868.
Jablonski, D., Jandzik, D., Mikulíček, P., Džukić, G., Ljubisavljević, K., Tzankov, N., Jelić, D., Thanou, E., Moravec, J., Gvoždík, V. (2016): Contrasting evolutionary histories of the legless lizards slow worms (Anguis) shaped by the topography of the Balkan Peninsula. BMC Evolutionary Biology 16: 99.
Jablonski, D., Najbar, B., Grochowalska, R., Gvoždík, V., Strzała, T. (2017): Phylogeography and postglacial colonization of Central Europe by Anguis fragilis and Anguis colchica. Amphibia-Reptilia 38: 562-569.
Jablonski, D., Sillero, N., Oskyrko, O., Bellati, A., Čeirāns, A., Cheylan, M., Cogălniceanu, D., Crnobrnja-Isailović, J., Crochet, P.-A., Crottini, A., Doronin, I., Džukić, G., Geniez, P., Ilgaz, C., Iosif, R., Jandzik, D., Jelić, D., Litvinchuk, S., Ljubisavljević, K., Lymberakis, P., Mikulíček, P., Mizsei, E., Moravec, J., Najbar, B., Pabijan, M., Pupins, M., Sourrouille, P., Strachinis, I., Szabolcs, M., Thanou, E., Tzoras, E., Vergilov, V., Vörös, J., Gvoždík, V. (2021): The distribution and biogeography of slow worms (Anguis, Squamata) across the Western Palearctic, with an emphasis on secondary contact zones. Amphibia-Reptilia 42: 519-530.
Kumar, S., Stecher, G., Li, M., Knyaz, C., Tamura, K. (2018): MEGA X: molecular evolutionary genetics analysis across computing platforms. Molecular Biology & Evolution 35: 1547-1549.
Lescure, J., de Massary, J.-C. (2012): Atlas des Amphibiens et Reptiles de France. Biotope, Mèze, Paris: MNHN. 272 pp.
Lui, J., Jiang, J., Song, S., Tornabene, L., Chabarria, R., Naylor, G.J.P., Li, C. (2017): Multilocus DNA barcoding - species identification with multilocus data. Scientific Reports 7: 16601.
Padial, J.M., Miralles, A., de la Riva, I., Vences, M. (2010): The integrative future of taxonomy. Frontiers in Zoology 7: 16.
Payseur, B.A. (2010): Using differential introgression in hybrid zones to identify genomic regions involved in speciation. Molecular Ecology Resources 10: 806-882.
Rasolonjatovo, S.M., Scherz, M.D., Hutter, C.R., Glaw, F., Rakotoarison, A., Razafindraibe, J.H., Goodman, S.M., Raselimanana, A.P., Vences, M. (2020): Sympatric lineages in the Mantidactylus ambreensis complex of Malagasy frogs originated allopatrically rather than by in-situ speciation. Molecular Phylogenetics & Evolution 144: 106700.
Renet, J., Lucente, D., Delaugerre, M., Gerriet, O., Deso, G., Abbattista, C., Cimmaruta, R. (2018): Discovery of an Italian slow worm (Anguis veronensis Pollini, 1818) population on a Western Mediterranean Island confirmed by genetic analysis. Acta Herpetologica 13: 165-169.
Rubinoff, D., Holland, B.S. (2005): Between two extremes: mitochondrial DNA is neither the panacea nor the nemesis of phylogenetic and taxonomic inferences. Systematic Biology 54: 952-961.
Rubinoff, D., Cameron, S., Will, K. (2006): A genomic perspective on the shortcomings of mitochondrial DNA for “barcoding” identification. Journal of Heredity 97: 581-594.
Sillero, N., Campos, J., Bonardi, A., Corti, C., Creemers, R., Crochet, P.A., Crnobrnja-Isailovic, J., Denoël, M., Ficetola, G.F., Gonçalves, J., Kuzmin, S., Lymberakis, P., de Pous, P., Rodriguez, A., Sindaco, R., Speybroeck, J., Toxopeus, B., Vieites, D.R., Vences, M. (2014): Updated distribution and biogeography of amphibians and reptiles of Europe. Amphibia Reptilia 25: 1-31.
Singhal, S., Moritz, C. (2013): Reproductive isolation between phylogeographic lineages scales with divergence. Proceedings of the Royal Society B: Biological Sciences 280: 20132246.
Speybroeck, J., Beukema, W., Dufresnes, C., Fritz, U., Jablonski, D., Lymberakis, P., Martínez-Solano, I., Razzetti, E., Vamberger, M., Vences, M., Vörös, J., Crochet, P.-A. (2020): Species list of the European herpetofauna - 2020 update by the Taxonomic Committee of the Societas Europaea Herpetologica. Amphibia-Reptilia 41: 139-189.
Szabó, K., Vörös, J. (2014): Distribution and hybridization of Anguis fragilis and A. colchica in Hungary. Amphibia-Reptilia 35: 135-140.
Thanou, E., Giokas, S., Kornilios, P. (2014): Phylogeography and genetic structure of the slow worms Anguis cephallonica and Anguis graeca (Squamata: Anguidae) from the southern Balkan Peninsula. Amphibia-Reptilia 35: 263-269.
Thanou, E., Kypraios-Skrekas, V., Kornilios, P., Giokas, S. (2021): Ecomorphological divergence and lack of gene flow in two sympatric Balkan slow worms (Squamata: Anguidae). Biological Journal of the Linnean Society 134: 443-460.
Ursenbacher, S., Zwahlen, V. (2015): Les orvets: une ou deux espèces en Suisse?. In: Résumés des communications présentées lors du 22ème Colloque herpétologique du Centre de Coordination pour la Protection des Amphibiens et des Reptiles de Suisse (karch). 5th December 2015; Bern, Switzerland.
Vences, M., Thomas, M., Bonett, R.M., Vieites, D.R. (2005): Deciphering amphibian diversity through DNA barcoding: chances and challenges. Philosophical Transactions of the Royal Society B: Biological Sciences 360: 1859-1868.
