title: A new PCR-RFLP-based method for an easier systematic affiliation of European water frogs.
abstract: We describe a non-invasive, PCR-RFLP-based method that allows reliable determination of the European water frog species Pelophylax lessonae and Pelophylax ridibundus and the hybrid form Pelophylax esculentus. Maximum-likelihood analysis of ITS2 sequences revealed two robust monophyletic clades corresponding to water frogs of the P. lessonae and P. ridibundus groups. Three restriction enzymes (KpnI, HaeII, and SmaI) were used to digest three conserved ITS2 domains. Taxonomic identification was unambiguous; the three restriction enzymes gave the same results. A French reference sample was identified using allozyme electrophoresis. Our PCR-RFLP method confirmed circa 83% of identification of the allozyme method. We conclude that the difference between identifications was caused by introgression.
authors.affiliation: LEESA, GECCO, Université d'Angers, Campus Belle Beille, 2 bvd Lavoisier, F-49045 Angers Cedex, France. cecile.patrelle@gmail.fr
authors.forename: Cécile
authors.initials: C
authors.lastname: Patrelle
authors.forename: Torsten
authors.initials: T
authors.lastname: Ohst
authors.forename: Damien
authors.initials: D
authors.lastname: Picard
authors.forename: Alain
authors.initials: A
authors.lastname: Pagano
authors.forename: Stéphane
authors.initials: S
authors.lastname: Sourice
authors.forename: Marie-Gaëlle
authors.initials: MG
authors.lastname: Dallay
authors.forename: Jörg
authors.initials: J
authors.lastname: Plötner
mesh_terms: D000818:Animals; D005060:Europe; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D011898:Ranidae / Q000145:classification* / Q000235:genetics*