Isolation and characterisation of 36 polymorphic microsatellite markers using 454 sequencing in the bar-shouldered skink
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TECHNICAL NOTE
Isolation and characterisation of 36 polymorphic microsatellite markers using 454 sequencing in the bar-shouldered skink, Ctenotus inornatus E. Harradine • R. A. How • L. H. Schmitt C. de Hass • P. B. S. Spencer
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Received: 24 August 2012 / Accepted: 27 August 2012 / Published online: 6 September 2012 Ó Springer Science+Business Media B.V. 2012
Abstract We report on the development of 36 novel polymorphic microsatellite markers from the bar-shouldered skink, Ctenotus inornatus. Members in this genus account for up to 20 % of the diverse Australian lizard species. Microsatellite markers were developed to investigate island endemism amongst Kimberley islands taxa. We describe the diversity from 110 adult Ctenotus individuals. Overall, the markers produced between 9 to 40 alleles per locus and observed and expected heterozygosities of 74.6 and 95.7 %, respectively. Due to island endemism, habitat modification by introduced grazing mammals, some urban encroachment and changes in fire regime there has been fragmentation of its range. These microsatellite markers are an important tool for evaluating population genetic information and for prioritising populations for conservation management. Keywords Conservation Ctenotus fallens Ctenotus inornatus Genetic diversity Microsatellite Fragmentation Scincidae Sphenomorphus group
E. Harradine P. B. S. Spencer (&) School of Biological Sciences and Biotechnology, Murdoch University, Perth, WA 6150, Australia e-mail: [email protected] R. A. How Department of Terrestrial Vertebrates, Western Australian Museum, Perth, WA, Australia L. H. Schmitt C. de Hass School of Anatomy and Human Biology, The University of Western Australia, Crawley, WA 6009, Australia
The genus Ctenotus is one of the most diverse lizard genera in Australia, with an estimated 100 species inhabiting most of Australia excluding Tasmania and high mountain tops (Cogger 2000; Rabosky et al. 2007). Their diversity, abundance and relative ease to sample have made Ctenotus a popular system to model a number of ecosystem and species attributes including species richness, diversification and sympatry (Rabosky et al. 2007). Ctenotus inornatus inhabit a wide range of habitat types throughout subhumid and semiarid northern Australia including many islands, making it an ideal species to examine the effects of insularity on island populations (Storr 1975). For this purpose we report on the development of polymorphic loci to provide sufficient resolution for detecting fine-scale population genetic structure, gene flow and measures of population genetic diversity. We also report on the use of 18 of these markers for Ctenotus fallens, a medium- sized skink found along coastal Western Australia between Perth and Shark Bay (Cogger 2000). Microsatellites were isolated using a second generation sequencing approach similar to that of Abdelkrim et al. (2009). Genomic DNA was isolated from liver (WA Museum voucher accession number R171779) using a using QIAGEN DNeasy tissue/blood kit (QIAGEN I
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