Characterisation of nine European wall lizard ( Podarcis muralis ) microsatellite loci of utility across sub-species
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MICROSATELLITE LETTERS
Characterisation of nine European wall lizard (Podarcis muralis) microsatellite loci of utility across sub-species Robert J. P. Heathcote • Deborah A. Dawson Tobias Uller
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Received: 24 July 2014 / Accepted: 29 July 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract Human-assisted dispersal of the European wall lizard (Podarcis muralis) has created conservation concerns due to extensive introgression between normally allopatric and genetically distinct lineages. We characterised nine microsatellite loci that amplified in four mtDNA clades belonging to two main evolutionary lineages (broadly belonging to P. m. brogniardii and P. m. nigriventris) known to hybridise in England and Germany. All loci were autosomal and displayed 5–28 alleles. These loci will prove useful for population genetic and parentage studies in hybrid zones. Keywords Hybridisation Introgression Paternity Podarcis muralis The European wall lizard (Podarcis muralis) consists of multiple lineages distributed across Europe. Human introductions of this species have created conservation concerns due to extensive hybridisation between genetically distinct Electronic supplementary material The online version of this article (doi:10.1007/s12686-014-0294-5) contains supplementary material, which is available to authorized users. R. J. P. Heathcote (&) T. Uller Edward Grey Institute, Department of Zoology, University of Oxford, South Parks Road, Oxford OX1 3PS, UK e-mail: [email protected] R. J. P. Heathcote D. A. Dawson Department of Animal and Plant Sciences, University of Sheffield, Sheffield S10 2TN, UK Present Address: R. J. P. Heathcote Centre for Research in Animal Behaviour, College of Life and Environmental Sciences, University of Exeter, Perry Road, Exeter EX4 4QJ, UK
and naturally allopatric lineages (Schulte et al. 2012). Since hybridisation is asymmetrical with respect to sex (Schulte et al. 2012), its study requires nuclear markers with crosslineage amplification. We isolated microsatellite markers in P. muralis and characterised them in four mtDNA clades; two from France (‘French’ lineage, broadly corresponding to P. m. brogniardii) and two from Italy (‘Italian’ lineage, broadly corresponding to P. m. nigriventris) (Michaelides et al. 2013). Genomic DNA was extracted from tail-tips using the ammonium acetate precipitation method (Nicholls et al. 2000). We created a microsatellite-enriched library using an Italian female from Ventnor, England (ID:497; Tuscan mtDNA clade) (following Mills et al. 2013). Twenty-two 454 microsatellite sequences were selected that were a consensus of multiple sequences. Their uniqueness was confirmed using BLASTN v.2.2.4 (Altschul et al. 1997). Primer sets were designed using Primer3v.0.4.0 and initially tested in 24 French animals caught from an introduced population at Cheyne Weare, Dorset, UK. Fifteen markers were discarded that were monomorphic, amplified non-specific products or failed to amplify in multiple individuals. Nine markers amplified reproduci
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