White breasted-mesites (Mesitornis variegata) are endemic birds from the dry deciduous forests of western Madagascar. They are monomorphic, ground-dwelling, medium-sized birds found in pairs or small groups (Hawkins and Seddon 2003). There is only little known on the general biology of this species, and its population genetic structure has not been studied. Here we describe the isolation and characterization of 10 microsatellite loci that were developed to study the social organization and mating system of M. variegata. White-breasted mesites are classified as vulnerable in the IUCN red list of threatened species (BirdLife International 2012). Available specific microsatellite markers can be an important tool for species conservation and could be used to assess the viability of the few remaining populations of this species by determining their genetic variability and degree of isolation (Hedrick 2001).

Feather, blood and tissue samples were collected in Kirindy Forest (Kappeler and Fichtel 2012), from birds and embryo remains of predated eggs. Sample collection and export were conducted according to local authority permits.

Microsatellite sequences were isolated by ecogenics GmbH (Switzerland). Size selected fragments from genomic DNA were enriched for simple sequence repeat (SSR) content by using magnetic streptavidin beads and biotin-labeled CT and GT repeat oligonucleotides. The SSR enriched library was analyzed on a Roche 454 platform using the GS FLX titanium reagents. The total 18,013 reads had an average length of 201 base pairs. Of these, 1,021 contained a microsatellite insert with a tetra- or a tri-nucleotide of at least 6 repeat units or a dinucleotide of at least 10 repeat units. Suitable primer design was possible in 340 reads, of which 36 were tested for polymorphism. We extracted DNA from 75 individuals using DNeasy Blood and Tissue Kit (Qiagen). Reactions of 10 μl containing 1× buffer, 200 μM of dNTPs, 0.04 μM M13 tailed locus specific forward primer, 0.16 μM locus specific reverse primer, 0.16 μM universal M13 primer 5′-end labeled with FAM (Metabion), 0.5 units of Hotstar Taq (Qiagen) were used to amplify each locus via the nested PCR procedure described by Schuelke (2000). The PCR profile was 95 °C for 15 min, 30 cycles of 30 s at 95 °C, 45 s at 56 °C (annealing temperature), 45 s at 72 °C, followed by 8 cycles of 30 s at 95 °C, 45 s at 53 °C and 45 s at 72 °C; and a final elongation phase of 30 min at 72 °C. PCR products were sized on a 3130XL Genetic Analyzer (Applied Biosystems/Hitachi) and GENEMAPPER V4 (Applied Biosystems) was used to assign genotypes. Observed (Ho) and expected (He) heterozygosity and Hardy–Weinberg equilibrium were calculated using GENEPOP V4.1.4 (Rousset 2008). Description of 10 polymorphic microsatellite loci is provided in Table 1. We detected 4–10 alleles per locus (mean = 6.1), Ho ranged from 0.288 to 0.853 and nine loci were in Hardy–Weinberg equilibrium. Overall, the described microsatellite markers should be an adequate tool for the study of the social system and conservation of M. variegata.

Table 1 Characterization of 10 microsatellite loci for M. variegata from 75 individuals
Full size table