[en] Water frogs (genus Pelophylax) are one of the most widespread and diverse, but also most invasive amphibians of the Western Palearctic region. As such, Pelophylax studies face the challenge of identifying similar taxa that hybridize in sympatry. For this purpose, the nuclear marker serum albumin intron 1 (SAI-1) has been used for over a decade in Pelophylax. Initially praised for its diagnosticity, notably to discriminate common species such as the pool frog (P. lessonae), the marsh frog (P. ridibundus) and their hybridogenetic hybrid the edible frog (P. esculentus) without sequencing (by amplicon length polymorphism), SAI-1 was later questioned due to misidentifications and doubtful patterns of genetic divergence. In this study, we incorporate an up-to-date multilocus phylogeographic framework spanning the entire Pelophylax diversification, to reassess the performance of SAI-1 for lineage identification and discovery. We show that SAI-1 sequences discriminate all Palearctic water frog species and most of their phylogeographic lineages, enabling us to map their distributions and identify the genomes of hybridogenetic hybrids. However, the phylogeny of SAI-1 is aberrant and unrepresentative of the evolution of the genus. In particular, differentiated P. l. lessonae alleles segregating in the Alpine region mimic a species-level divergence that is not recovered by any other marker. Moreover, the indel polymorphism that supposedly distinguishes P. lessonae from P. ridibundus, as well as the main P. ridibundus lineages from the Balkans (P. r. ridibundus vs kurtmuelleri), are not diagnostic across the entire range of these taxa. Hence, SAI-1 is neither the panacea for nor the nemesis of Pelophylax genotyping. Sequencing SAI-1 shall continue to offer a reliable and informative preliminary approach of single-gene barcoding identification of lineages, but analyses without sequencing, and other applications such as phylogenetic and taxonomic inferences, should be avoided.
Research Center/Unit :
FOCUS - Freshwater and OCeanic science Unit of reSearch - ULiège
Disciplines :
Genetics & genetic processes
Author, co-author :
Dufresnes, Christophe; Institut de Systématique, Muséum national d'Histoire naturelle, Evolution ; CNRS, Sorbonne Université, EPHE, Université des Antilles, Paris, France
Dubey, Sylvain; Department of Ecology and Evolution, University of Lausanne, Lausanne, Switzerland ; HW Romandie SA
Wielstra, Ben; Institute of Biology, Leiden University, Leiden, The Netherlands ; Naturalis Biodiversity Center, Leiden, The Netherlands
Jablonski, Daniel; Department of Zoology, Comenius University in Bratislava, Bratislava, Slovakia
Denoël, Mathieu ; Université de Liège - ULiège > Freshwater and OCeanic science Unit of reSearch (FOCUS) ; Université de Liège - ULiège > Département de Biologie, Ecologie et Evolution > Laboratoire d'Écologie et de Conservation des Amphibiens (LECA)
Language :
English
Title :
Panacea or nemesis? Re-assessing the reliability of serum albumin intron 1 for genotyping Western Palearctic water frogs
Publication date :
2024
Journal title :
Alytes
ISSN :
0753-4973
Publisher :
International Society for the Study and Conservation of Amphibians, France
Arikan, H. (1988) On a new form of Rana ridibunda (Anura, Ranidae) from Turkey. İstanbul Üniversitesi fen Fakültesi Biyoloji Dergisi, 53: 81–87.
Beerli, P., Hotz, H., Tunner, H. G., Heppich, S. & Uzzell, T. (1994) Two new water frog species from the Aegean Islands Crete and Karpathos (Amphibia, Salientia, Ranidae). Notulae Naturae of the Academy of natural Sciences of Philadelphia, 470: 1–9.
Bonaparte, C. L. J. L. (1839) Iconografia della Fauna Italica per le quattro classi degli Animali Vertebrati. Tomo II. Amphibi. Fascicolo 24. Rome (Salviucci): 1–560, 54 pl.
Camerano, L. (1882) Recherches sur les variations de la Rana esculenta et du Bufo viridis dans le Bassin de la Méditerranée. Comptes rendus de l’Association française pour l’Avancement des Sciences. Paris, 10: 680–690.
Christiansen, D. G. (2005) A microsatellite-based method for genotyping diploid and triploid water frogs of the Rana esculenta hybrid complex. Molecular Ecology Notes, 5 (1): 190–193.
Cuevas, A., Patrelle, C., Ciavatti, F., Gendre, T., Sourrouille, P., Geniez, P., Doniol-Valcroze, P. & Crochet, P.-A. (2022) A new PCR-RFLP method for the identification of parental and hybridogenetic western European water frogs, including the Pelophylax perezi-grafi system. Salamandra, 58 (3): 218–230.
Doniol-Valcroze, P., Mazepa, G., Grimal, F., Sourrouille, P., Perrin, N., Litvinchuk, S. N. & Crochet, P.-A. (2021) Discovery of a Pelophylax saharicus (Anura, Ranidae) population in southern France: a new potentially invasive species of water frogs in Europe. Amphibia-Reptilia, 42 (4): 427–442.
Dubey, S. & Dufresnes, C. (2017) An extinct vertebrate preserved by its living hybridogenetic descendant. Scientific Reports, 7 [12768]: 1–6.
Dubey, S., Maddalena, T., Bonny, L., Jeffries, D. L. & Dufresnes, C. (2019) Population genomics of an exceptional hybridogenetic system of Pelophylax water frogs. BMC evolutionary Biology, 19 [164]: 1–13.
Dubois, A. (1983) A propos de cuisses de grenouilles. Alytes, 2 (3): 69–111.
Dufresnes, C. (2024) Data from: Piecing the barcoding puzzle of Palearctic water frogs (Pelophylax) sheds light on amphibian biogeography and global invasions. Zenodo. .
Dufresnes, C., Denoël, M., Di Santo, L. & Dubey, S. (2017) Multiple uprising invasions of Pelophylax water frog, potentially inducing a new hybridogenetic complex. Scientific Reports, 7 [6506]: 1–10.
Dufresnes, C. & Dubey, S. (2020) Invasion genomics supports an old hybrid swarm of pool frogs in western Europe. Biological Invasions, 22 (2): 205–210.
Dufresnes, C. & Jablonski, D. (2022) A genomics revolution in amphibian taxonomy. Science, 377 (6612): 1272.
Dufresnes, C. & Mazepa, G. (2020) Hybridogenesis in water frogs. eLS, 1 (4): 718–726.
Dufresnes, C., Mazepa, G., Jablonski, D., Caliari Oliveira, R., Wenseleers, T., Shabanov, D. A., Auer, M., Ernst, R., Ramírez-Chaves, H. E., Mulder, K. P., Simonov, E., Tiutenko, A., Kryvokhyzha, D., Wennekes, P. L., Zinenko, O. I., Korshunov, O. V., Al-Johany, A. M., Peregontsev, E. A., Betto-Colliard, C., Denoël, M., Borkin, L. J., Skorinov, D. V., Pasynkova, R. A., Mazanaeva, L. F., Rosanov, J. M., Dubey, S. & Litvinchuk, S. N. (2019) Fifteen shades of green: the evolution of Bufotes toads revisited. Molecular Phylogenetics and Evolution, 141 [106615]: 1–25.
Dufresnes, C., Monod-Broca, M., Bellati, A., Canestrelli, D., Ambu, J., Wielstra, B., Dubey, S., Crochet, P.-A., Denoël, M. & Jablonski, D. (2024) Piecing the barcoding puzzle of Palearctic water frogs (Pelophylax) sheds light on amphibian biogeography and global invasions. Global Change Biology, 30 (3) [e17180]: 1–30.
Engelmann, W.-E., Fritzsche J., Günther, R. & Obst, F. J. (1986) Lurche und Kriechtiere Europas. Stuttgart (Ferdinand Enke): 1–420.
Ermakov, O., Ivanov, A., Titov, S., Svinin, A. & Litvinchuk, S. N. (2019) New multiplex PCR method for identification of East European green frog species and their hybrids. Russian Journal of Herpetology, 26 (6): 367–370.
Fitzinger, L. J. F. J. (1843). Systema Reptilium. Fasciculus Primus. Wien (Braumüller et Seidel): 1–106.
Gayda, H. S. (1940). Su alcuni anfibi e rettili dell’Albania esistenti nel Museo zoologico di Berlino. Atti della Societa italiana di Scienze naturali e del Museo civico di Storia naturale di Milano, 79 (4): 263–272.
Hallowell, E. (1861) Report upon the Reptilia of the North Pacific Exploring Expedition, under command of Capt. John Rogers, U.S. N. Proceedings of the Academy of natural Sciences of Philadelphia, 1860: 480–510.
Hartert, E. (1913) Expedition to the central western Sahara. V. Reptiles and Batrachians. Novitates zoologicae, 20: 76–84.
Hauswaldt, J. S., Höer, M., Ogielska, M., Christiansen, D. G., Dziewulska-Szwajkowska, D., Czernicka, E. & Vences, M. (2012) A simplified molecular method for distinguishing among species and ploidy levels in European water frogs (Pelophylax). Molecular Ecology Resources, 12 (5): 797–805.
Herczeg, D., Vörös, J., Christiansen, D. G., Benovics, M. & Mikulíček, P. (2017) Taxonomic composition and ploidy level among European water frogs (Anura: Ranidae: Pelophylax) in eastern Hungary. Journal of zoological Systematics and evolutionary Research, 55 (2): 129–137.
Hotz, H., Uzzell, T., Günther, R., Tunner, H. G. & Heppich, S. (1987) Rana shqiperica, a new European water frog species from the Adriatic Balkans (Amphibia, Salientia, Ranidae). Notulae Naturae of the Academy of natural Sciences of Philadelphia, 468: 1–3.
Hotz, H., Beerli, P., Uzzell, T., Guex, G.-D., Pruvost, N. B. M., Schreiber, R. & Plötner, J. (2013) Balancing a cline by influx of migrants: a genetic transition in water frogs of eastern Greece. Journal of Heredity, 104 (1): 57–71.
Huson, D. H. & Bryant, D. (2006) Application of phylogenetic networks in evolutionary studies. Molecular Biology and Evolution, 23 (2): 254–267.
Jelić, M., Franjević, D., Đikić, D., Korlević, P., Vucić, M., Jelić, D., Becking, T., Grandjean, F. & Klobučar, G. (2022) Hybrid swarm as a result of hybridization between two alien and two native water frog species (genus Pelophylax) in central Croatia: Crna Mlaka fishpond acting as a species melting pot? Biological Invasions, 24 (10): 3291–3304.
Kolenda, K., Pietras-Lebioda, A., Hofman, S., Ogielska, M. & Pabijan, M. (2017) Preliminary genetic data suggest the occurrence of the Balkan water frog, Pelophylax kurtmuelleri, in southwestern Poland. Amphibia-Reptilia, 38 (2): 187–196.
Linnaeus, C. (1758) Systema Naturae per regna tria naturae, secundum classes, ordines, genera, species, cum characteribus, differentiis, synonymis, locis. Editio decima, reformata. Tomus I. Holmiae (Laurentii Salvii): [i–iv] + 1–824.
Litvinchuk, S. N., Ivanov, A. Y., Lukonina, S. A. & Ermakov, O. A. (2020) A record of alien Pelophylax species and widespread mitochondrial DNA transfer in Kaliningradskaya Oblast’ (the Baltic coast, Russia). BioInvasions Records, 9 (3): 599–617.
López-Seoane, V. (1885) On two new forms of Rana from N.W. Spain. The Zoologist: a monthly Journal of natural History, (3), 9 (101): 169–172.
Mayer, M., Hawlitscheck, O., Zahn, A. & Glaw, F. (2013) Composition of twenty green frog populations (Pelophylax) across Bavaria, Germany. Salamandra, 49 (1): 31–44.
Miura, I., Vershinin, V., Vershinina, S., Lebedinskii, A., Trofimov, A., Sitnikov, I. & Ito, M. (2021) Hybridogenesis in the water frogs from western Russian territory: intrapopulation variation in genome elimination. Genes, 12 (2): 244.
Neveu, A. (1989) Rana ridibunda. In: J. Castanet & R. Guyétant (ed.), Atlas de répartition des amphibiens et reptiles de France, Paris (Société Herpétologique de France): 90–91.
Pallas, P. S. (1771) Reise durch verschiedene Provinzen des Russischen Reichs. Theil 1. St. Pétersbourg (Kayserlichen Academie der Wissenschaften): 1–504.
Papežík, P., Kubala, M., Jablonski, D., Doležálková-Kaštánková, M., Choleva, L., Benovics, M. & Mikulíček, P. (2021) Morphological differentiation of endemic water frogs (Ranidae: Pelophylax) from the southwestern Balkans. Salamandra, 57 (1): 105–123.
Papežík, P., Sciberras, A., Benovics, M., Sciberras, J., Deidun, A. & Mikulíček, P. (2024) Far from home: tracing the origin of non-native water frogs (genus Pelophylax) in Malta by molecular markers. Biological Invasions, 26 (4): 1045–1059.
Patrelle, C. Ohst, T., Picard, D., Pagano, A., Sourice, S., Dallay, M.-G. & Plötner, J. (2011) A new PCR-RFLP-based method for an easier systematic affiliation of European water frogs. Molecular Ecology Resources, 11 (1): 200–205.
Plötner, J., Akın Pekşen, Ç., Baier, F., Uzzell, T. & Can Bilgin, C. (2015) Genetic evidence for human-mediated introduction of Anatolian water frogs (Pelophylax cf. bedriagae) to Cyprus (Amphibia: Ranidae). Zoology in the Middle East, 61 (2): 125–132.
Plötner, J., Baier, F., Akın, C., Mazepa, G., Schreiber, R., Beerli, P., Litvinchuk, S. N., Bilgin, C. C., Borkin, L. & Uzzell, T. (2012) Genetic data reveal that water frogs of Cyprus (genus Pelophylax) are an endemic species of Messinian origin. Zoosystematics and Evolution, 88 (2): 261–283.
Plötner, J., Köhler, F., Uzzell, T., Beerli, P., Schreiber, R., Guex, G. D. & Hotz, H. (2009) Evolution of serum albumin intron-1 is shaped by a 5’ truncated non-long terminal repeat retrotransposon in western Palearctic water frogs (Neobatrachia). Molecular Phylogenetics and Evolution, 53 (3): 784–791.
Plötner, J., Uzzell, T., Beerli, P., Spolsky, C., Ohst, T., Litvinchuk, S. N., Guey, G.-D., Reyer, H.-U. & Hotz, H. (2008) Widespread unidirectional transfer of mitochondrial DNA: a case in western Palaearctic water frogs. Journal of evolutionary Biology, 21 (3): 668–681.
Puky, M., Schad, P. & Szövénji, G. (2005) Magyaroszag herpetologiai atlasza [Herpetological atlas of Hungary]. Budapest (Varangy Akciocsoport Egyesület): 1–207, 16 pl.
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.
Sagonas, K., Karameta, E., Kotsakiozi, P. & Poulakakis, N. (2020) Cross-species testing of nuclear markers in Pelophylax water frogs in Greece and examination of their power to detect genetic admixture. Amphibia-Reptilia, 41 (2): 253–259.
Schneider, H., Sofianidou T. S. & Kyriakopoulou-Sklavounou P. (1984) Bioacoustic and morphometric studies in water frogs (genus Rana) of Lake Ioannina in Greece, and description of a new species (Anura, Amphibia). Journal of zoological Systematics and evolutionary Research, 22 (4): 349–366.
Schneider, J. G. (1799) Historia Amphibiorum naturalis et literarariae. Fasciculus primus. Continens Ranas, Calamitas, Bufones, Salamandras et Hydros in genera et species descriptos notisque suis distinctos. Jena (Friederici Frommanni): 1–264.
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 (2): 139–189.
Trifinopoulos, J., Nguyen, L. T., Haeseler, A. von & Minh, B. Q. (2016) W-IQ-TREE: a fast online phylogenetic tool for maximum likelihood analysis. Nucleic Acids Research, 44 (W1): W232–235.
Vucić, M., Jelić, D., Klobučar, G. I., Prkljačić, B. & Jelić, M. (2018) Molecular identification of species and hybrids of water frogs (genus Pelophylax) from Lake Skadar, Southeast Adriatic drainages (Amphibia: Ranidae). Salamandra, 54 (2): 147–157.
Wielstra, B., Burke, T., Butlin, R. K. & Arntzen, J. W. (2017) A signature of dynamic biogeography: enclaves indicate past species replacement. Proceedings of the royal Society of London, (B), 284 (1868) [20172014]: 1–6.
Zieliński, P., Nadachowska-Brzyska, K., Wielstra, B., Szkotak, R., Covaciu-Marcov, S., Cogălniceanu, D. & Babik, W. (2013) No evidence for nuclear introgression despite complete mtDNA replacement in the Carpathian newt (Lissotriton montandoni). Molecular Ecology, 22 (7): 1884–1903.
Zimić, A., Berina, V., Šunje, E., Ćurić, A. & Kalamujic Stroil, B. (2020) Molecular identification of green frogs (Anura: Ranidae: Pelophylax) of western Balkans (Bosnia & Herzegovina and Montenegro). In: D. Jöjic (ed.), IV Symposium of biologists and ecologists of Republic of Srpska with international participation – SBERS2020, Banja Luka (Faculty of Natural Sciences and Mathematics, University of Banja Luka): 152.