Challenges of microsatellite development in Lepidoptera: Euphydryas aurinia(Nymphalidae) as a case study

E-mail Address: melthide.sinama@gmail.com; vincent.dubut@univ-provence.fr; caroline.costedoat@univ-provence.fr; andre.gilles@univ-provence.fr; marius.junker@gmx.de; thibaut.malausa@sophia.inra.fr; martinjf@supagro.inra.fr; gabriel.neve@univ-provence.fr; nicolas.pech@univ-provence.fr; thsh@uni-trier.de; marie.zimmermann@univ-tours.fr; emese.meglecz@univ-provence.frInternational audienceCurrently it remains difficult to obtain robust microsatellite markers for Lepidoptera. In an attempt to overcome the problemsassociated with developing microsatellite markers for this insect order we combined (i) biotin-enrichment protocol, (ii) nextgeneration pyrosequencing (through 454 GS-FLX Titanium technology) and (iii) the use of individuals collected from eight geographicallydistant European populations representing three subspecies of Euphydryas aurinia. Out of 96 stringently designed primerpairs, 12 polymorphic microsatellite loci amplified without obvious evidence of null alleles in eight individuals from different subspecies.Between five and seven of these loci showed full within population applicability and three revealed to be robust and transferablebetween populations and sub-species, providing a first step towards the development of a valuable and robust tool forstudying conservation issues and evolution in E. aurinia populations. Nevertheless, as in most studies dealing with Lepidopteramicrosatellites, null alleles were detected in most of the developed markers. Our results emphasize the need for further research inorder to better understand the complex evolution and organization of Lepidopteran genomes