Latonia nigriventer ; Mediterranean painted frogs (genus Discoglossus Otth, 1837) are distributed across western Europe, North Africa and some Mediterranean islands. Previous studies have focused on their phylogenetic relationships, but the taxonomic position of the Iberian taxa (D. galganoi and D. jeanneae) is still under debate. By using microsatellites, patterns and rates of admixture can be quantified. We report the characterisation of eighteen novel polymorphic microsatellite loci in Iberian painted frogs. These loci were also tested in all other species of Discoglossus and in the recently rediscovered and highly endangered relative Latonia nigriventer. Two to eleven loci amplified in these species, and the number of polymorphic loci ranged from zero (in Latonia) to eight (in D. scovazzi). The new markers will be useful in addressing questions related to the evolutionary history, population structure, and conservation of Iberian Discoglossus. They also have potential for use in the North African species D. scovazzi and D. pictus, the latter of which is an invasive species rapidly expanding its range in southeast France and northeast Spain. ; This research was funded by grants CGL2008-04271-C02-01/BOS and CGL2011-28300 (Ministerio de Ciencia e Innovación, Ministerio de Economía y Competitividad, Spain, and FEDER) and PPII10-0097- 4200 (Junta de Comunidades de Castilla La Mancha and FEDER) No IMS. JGR is supported by the Consejo Superior de Investigaciones Cientificas of Spain (CSIC) and the European Social Fund (ESF) (JAE-pre PhD fellowship); IMS is funded by Project >Biodiversidad y, Ecology and Global Change> co-financed by North Portugal Regional Operational Programme 2007/2013 (ON.2-O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF) and DS is supported by a post-doctoral grant (SFRH/BPD/66592/2009) funded by Fundação para a Ciência e Tecnologia (FCT, Portugal) under the Programa Operacional Potencial Humano - Quadro de Referência Estratégico Nacional funds from the European Social Fund and Portuguese Ministério da Educação e Ciência. ; Peer Reviewed