Isolation and characterisation of the first microsatellite markers for the European stag beetle, Lucanus cervus (Coleoptera: Lucanidae)

The European stag beetle, Lucanus cervus, is recognised as a flagship species for biodiversity conservation. Although the species is widely distributed across Europe declines have led to it being granted protected or endangered status in a number of countries and regarded as "near threatened" by the IUCN. The integration of genetic approaches into conservation efforts is urgently needed but has been impeded to date by the lack of appropriate genetic markers. To provide such a resource the development of the first microsatellite loci for stag beetle is described. Loci were identified using two methods (i) enriched library cloning (ELC) and (ii) Restriction enzyme Associated DNA sequencing (RAD-Seq). Inefficient microsatellite detection using the ELC method suggests that RAD-Seq, or other Next Generation Sequencing based methods, may ultimately be more cost effective for obtaining informative suites of markers for this and other coleopteran species. 18 loci were characterised by genotyping 42 UK specimens collected as prey leftover/roadkill. All loci produced unambiguous genotypes and were polymorphic. Though preliminary, estimates of genetic variability suggest UK populations may be genetically depauperate. The microsatellite loci represent a suite of genetic markers that can be applied to non-invasive population monitoring and numerous other areas of Lucanus conservation and evolutionary research.