Microsatellite markers for the Iberian endemic Bosca’s newt, Lissotriton boscai (Caudata, Salamandridae)

Abstract

The new generation Roche/454 sequencing of DNA fragments enriched for microsatellite loci was used to isolated microsatellite markers for the Iberian endemic Bosca’s newt, Lissotriton boscai. Two multiplex PCR sets were optimized in order to genotype ten polymorphic tetranucleotide microsatellite loci. The level of genetic diversity of these loci was assessed in 42 individuals from three central-south populations of Portugal (Nazare, Alcobaca and Evora). The number of alleles per locus ranged from 11 to 20 (mean; Na = 16). Observed and expected heterozygosities ranged from 0.50 to 1.0 and 0.55 to 0.92, respectively. We found no deviations from Hardy–Weinberg equilibrium, nor did we find linkage disequilibrium between pairs of loci after Bonferroni correction. We found no evidence for large allele dropouts or stuttering, although null alleles were detected for loci Ltb17 and Ltb4 in Alcobaca population. These markers will be useful for resolving fine-scale population genetic structure, especially in contact zones between highly divergent lineages of L. boscai.