Abstract
The Western Spadefoot, Pelobates cultripes (Anura, Pelobatidae), is endemic to the Iberian Peninsula and southeastern France, with isolated populations in the Atlantic coast of France. Its populations are fragmented and it is considered Near Threatened by the IUCN. Here we describe the development of sixteen polymorphic microsatellite loci in this species. Polymorphism was assessed in 95 individuals from five Iberian populations. The number of alleles and expected heterozygosity ranged from 3 to 14 and 0.20 to 0.76, respectively. Eight loci cross-amplified in the closely related and Endangered Moroccan Spadefoot toad, Pelobates varaldii. These markers will be useful to address questions about the ecology, population genetics and evolutionary history of P. cultripes, including information on effective population size, habitat use and dispersal patterns, which are essential for the efficient management of the fragmented populations characteristic of most of its range.
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References
Andrés JA, Bogdanowicz SM (2011) Isolating microsatellite loci: looking back, looking ahead. In: Orgogozo V, Rockman MV (eds) Molecular methods for evolutionary genetics. Springer, New York (USA), pp 211–232
Araújo MB, Thuiller W, Pearson RG (2006) Climate warming and the decline of amphibians and reptiles in Europe. J Biogeogr 33:1712–1728
Beja P, Bosch J, Tejedo M, Lizana M, Martínez-Solano I, Salvador A, García-París M, Recuero Gil E, Pérez-Mellado V, Díaz Paniagua C, Cheylan M, Márquez R, Geniez P (2009) Pelobates cultripes. In: IUCN 2012. IUCN Red List of Threatened Species. Version 2012.2. http://www.iucnredlist.org. Accessed 10 April 2013
Belkhir K, Borsa P, Chikhi L, Raufaste N, Bonhomme F (2000) GENETIX 4.04, Logiciel sous Windows TM pour la Génétique des Populations. Laboratoire Génome, Populations, Interactions. CNRS UMR 5000, Université de Montpellier II, Montpellier, France
Duget R, Melki F (eds) (2003) Les Amphibiens de France, Belgique et Luxemburg, Collection Parthénope. Editions Biotope, Mèze (France)
García-París M, Montori A, Herrero P (2004) Fauna Iberica. Vol. 24. Amphibia: Lissamphibia. Museo Nacional de Ciencias Naturales, Consejo Superior de Investigaciones Científicas, Madrid (Spain)
Hof C, Araújo MB, Jetz W, Rahbek C (2011) Additive threats from pathogens, climate and land-use change for global amphibian diversity. Nature 480:516–519
Holleley CE, Geerts PG (2009) Multiplex Manager 1.0: a cross-platform computer program that plans and optimizes multiplex PCR. Biotechniques 46:511–517
IUCN, Conservation International, NatureServe (2008) An analysis of amphibians on the 2008 IUCN Red List. http://www.iucnredlist.org/amphibians. Accessed 6 October 2008
Loureiro A, Carretero MA, Ferrand N, Paulo O (2008) Atlas dos Anfíbios e Répteis de Portugal Continental. Instituto da Conservação da Natureza, Lisboa (Portugal)
Meglécz E, Costedoat C, Dubut V et al (2010) QDD: a user-friendly program to select microsatellite markers and design primers from large sequencing projects. Bioinformatics 26:403–404
Peakall R, Smouse PE (2006) GenAlEx 6: genetic analysis in Excel. Population genetic software for teaching and research. Mol Ecol Notes 6:288–295
Rice WR (1989) Analyzing tables of statistical tests. Evolution 43:223–225
Rousset F (2008) Genepop’007: a complete re-implementation of the genepop software for Windows and Linux. Mol Ecol Resour 8:103–106
Salvador A, Donaire-Barroso D, Slimani T, El Mouden EH, Geniez P (2004) Pelobates varaldii. In: IUCN 2012. IUCN Red List of Threatened Species. Version 2012.2. http://www.iucnredlist.org. Accessed 19 April 2013
Stuart SN, Hoffmann M, Chanson JS, Cox NA, Berridge RJ, Ramani P, Young BE (2008) Threatened amphibians of the world. Lynx Edicions, Barcelona, Spain; IUCN, Gland, Switzerland; and Conservation International, Arlington, VA, USA
Tejedo M, Reques R (2002) Pelobates cultripes (Cuvier, 1829). Sapo de espuelas. In: Pleguezuelos JM, Márquez R, Lizana M (eds) Atlas y Libro Rojo de los Anfibios y Reptiles de España. Dirección General de Conservación de la Naturaleza-Asociación Herpetológica Española, Madrid (Spain), pp 94–96
Van de Vliet M, Diekmann O, Serrão E, Beja P (2009) Development and characterization of highly polymorphic microsatellite loci for the Western Spadefoot toad, Pelobates cultripes. Conserv Genet 10:993–996
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 4:535–538
Acknowledgments
We thank M. Barbosa, D. Buckley, I. Gómez Mestre, E. Recuero, G. Sánchez and V. Sancho for help collecting samples and S. Bogdanowicz at Cornell University for help with the microsatellite library. 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) to IMS. JGR is supported by the Consejo Superior de Investigaciones Científicas of Spain (CSIC) and the European Social Fund (ESF) (JAE-pre PhD fellowship), and IMS is a ‘Ramón y Cajal’ postdoctoral fellow supported by the Spanish Ministerio de Ciencia e Innovación and the Universidad de Castilla la Mancha.
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Gutiérrez-Rodríguez, J., Martínez-Solano, I. Isolation and characterization of sixteen polymorphic microsatellite loci in the Western Spadefoot, Pelobates cultripes (Anura: Pelobatidae) via 454 pyrosequencing. Conservation Genet Resour 5, 981–984 (2013). https://doi.org/10.1007/s12686-013-9948-y
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DOI: https://doi.org/10.1007/s12686-013-9948-y