Conservation Genetics Resources

, Volume 3, Issue 2, pp 311–313 | Cite as

Thirteen polymorphic microsatellite markers for the European green toad Bufo viridis viridis, a declining amphibian species

  • Christophe Dufresnes
  • Caroline Betto-Colliard
  • Nicolas Perrin
  • Matthias Stöck
Technical Note


We report 13 new polymorphic microsatellite markers for the European green toad Bufo viridis viridis (B. viridis subgroup), a declining amphibian from Central, Southeastern and Eastern Europe. Diversity at these loci estimated for 19 individuals ranged from two to ten alleles. Most of these primers also cross-amplify in related West-Mediterranean green toad species (Bufo balearicus, B. siculus and B. boulengeri). These microsatellites will be useful for conservation genetics of threatened Bufo viridis viridis populations and evolutionary studies of green toad taxa in secondary contact to examine hybridization.


Bufo viridis viridis Polymorphic microsatellites Population genetics Conservation research 



We thank Lucio Bonato, Nicola Novarini, Stefano Mazzotti, and G. Francesco Ficetola for providing samples that served, in part, to choose pure B. viridis viridis populations examined in this paper. This study was in part supported by a grant from the “Fondation 450ème anniversaire” of the University of Lausanne (November 23, 2009 to M.St.).


  1. Berne Convention (1979) Bern convention on the conservation of European wildlife and natural habitats 1979, also known as the Bern convention, came into force on 1 June 1982, (
  2. Colliard C, Sicilia A, Moritz C, Perrin N, Stöck M (2009) New polymorphic microsatellite markers and development of mitotyping primers for West Mediterranean green toad species (Bufo viridis subgroup). Mol Ecol Resour 9(4):1138–1140CrossRefGoogle Scholar
  3. Colliard C, Sicilia A, Turrisi GF, Arculeo M, Perrin N, Stöck M (2010) Strong reproductive barriers in a narrow hybrid zone of West-Mediterranean green toads (Bufo viridis subgroup) with Plio-Pleistocene divergence. BMC Evol Biol 10:232CrossRefPubMedGoogle Scholar
  4. Excoffier L, Laval G, Schneider S (2005) Arlequin (version 3.0): an integrated software package for population genetics data analysis. Evol Bioinform 1:47–50Google Scholar
  5. Goudet J (1995) Fstat (version 1.2): a computer program to calculate F-statistics. J Hered 86:485–486; program available:
  6. Grossenbacher K (2003) Die Wechselkröte (Bufo viridis) in der Schweiz: historische und aktuelle Situation. Mertensiella 14:147–152Google Scholar
  7. Habitats directive (1992) Habitats directive—council directive 92/43/EEC on the conservation of natural habitats and of wild fauna and flora.
  8. Kilpatrick C, Briggs J, Daszak P (2010) The ecology and impact of chytridiomycosis: an emerging disease of amphibians. Trends Ecol Evol 25(2):109–118CrossRefPubMedGoogle Scholar
  9. Plan National (2010) Plan national d’action crapaud vert (pour La France). DREAL Lorraine—BIOTOPE Draft: Août 2010; see:
  10. Podloucky R, Manzke U (2003) Verbreitung, Ökologie und Schutz der Wechselkröte (Bufo viridis). DGHT Rheinbach, Mertensiella 14:1–327Google Scholar
  11. Rowe G, Beebee T, Burke T (2000) A further four polymorphic microsatellite loci in the natterjack toad Bufo calamita. Conserv Genet 1:371–372CrossRefGoogle Scholar
  12. Stöck M, Moritz C, Hickerson M, Frynta D, Dujsebayeva T, Eremchenko V, Macey JR, Papenfuss TJ, Wake DB (2006) Evolution of mitochondrial relationships and biogeography of Palearctic green toads (Bufo viridis subgroup) with insights in their genomic plasticity. Mol Phylogenet Evol 41:663–689CrossRefPubMedGoogle Scholar
  13. Stöck M, Roth P, Podloucky R, Grossenbacher K (2008) Wechselkröten – unter Berücksichtigung von Bufo viridis virdis Laurenti, 1768; Bufo variabilis (Pallas, 1769); Bufo boulengeri Lataste, 1879; Bufo balearicus Böttger, 1880 und Bufo siculus Stöck, Sicilia, Belfiore, Lo Brutto, Lo Valvo und Arculeo, 2008. In: Grossenbacher K (ed) Handbuch der Amphibien und Reptilien Europas. vol. 5 (Froschlurche II) [Handbook of the Amphibians and Reptiles of Europe. vol. 5 (Anura II)]. AULA-Verlag, Wiesbaden (invited book chapter), pp. 413–498Google Scholar
  14. Stöck M, Ustinova J, Lamatsch D, Schartl M, Perrin N, Moritz C (2010) A vertebrate reproductive system involving three ploidy levels: hybrid origin of triploids in a contact zone of diploid and tetraploid Palearctic green toads (Bufo viridis subgroup). Evolution 64(4):944–959Google Scholar
  15. Van Oosterhout C, Hutchinson WF, Wills DPM, Shipley P (2004) MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data. Mol Ecol Notes 4:535–538CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Christophe Dufresnes
    • 1
  • Caroline Betto-Colliard
    • 1
  • Nicolas Perrin
    • 1
  • Matthias Stöck
    • 1
  1. 1.Department of Ecology and EvolutionUniversity of LausanneLausanneSwitzerland

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