Microsatellite variability is widely used to infer levels of genetic diversity in natural populations. However, the ascertainment bias caused by typically selecting only the most polymorphic markers in the genome may lead to reduced sensitivity for judging genome-wide levels of genetic diversity. To test this potential limitation of microsatellite-based approaches, we assessed the degree of nucleotide diversity in noncoding regions of eight different carnivore populations, including inbred as well as outbred populations, by sequencing 10 introns (5.4-5.7 kb) in 20 individuals of each population (wolves, coyotes, wolverines and lynxes). Estimates of nucleotide diversity varied 30-fold (7.1 x 10(-5 )-2.1 x 10(-3)), with densities of one single nucleotide polymorphism every 112-5446 bp. Microsatellite genotyping (10-27 markers) of the same animals revealed mean multilocus heterozygosities of 0.54-0.78, a 1.4-fold difference among populations. There was a positive yet not perfect (r(2) = 0.70) correlation between microsatellite marker heterozygosity and nucleotide diversity at the population level. For example, point estimates of nucleotide diversity varied in some cases with an order of magnitude despite very similar levels of microsatellite marker heterozygosity. Moreover, at the individual level, no significant correlation was found. Our results imply that variability at microsatellite marker sets typically used in population studies may not accurately reflect the underlying genomic diversity. This suggests that researchers should consider using resequencing-based approaches for assessing genetic diversity when accurate inference is critical, as in many conservation and management contexts.

Background: We introduce the use of short insertion-deletion polymorphisms (indels) for genetic analysis of natural populations.

Hybridization is a significant threat for endangered species and could potentially even lead to their extinction. This concern applies to the globally vulnerable Greater Spotted Eagle Aquila clanga, a species that co-occurs, and potentially interbreeds, with the more common Lesser Spotted Eagle Aquila pomarina in a vast area of Eastern Europe. We applied single nucleotide polymorphism (SNP) and microsatellite markers in order to study hybridization and introgression in 14 European spotted eagle populations. We detected hybridization and/or introgression in all studied sympatric populations. In most regions, hybridization took place prevalently between A. pomarina males and A. clanga females, with introgression to the more common A. pomarina. However, such a pattern was not as obvious in regions where A. clanga is still numerous. In the course of 16 years of genetic monitoring of a mixed population in Estonia, we observed the abandonment of A. clanga breeding territories and the replacement of A. clanga pairs by A. pomarina, whereby on several occasions hybridization was an intermediate step before the disappearance of A. clanga. Although the total number of Estonian A. clanga ¥ A. pomarina pairs was twice as high as that of A. clanga pairs, the number of pairs recorded yearly were approximately equal, which suggests a higher turnover rate in interbreeding pairs. This study shows that interspecific introgressive hybridization occurs rather frequently in a hybrid zone at least 1700-km wide: it poses an additional threat for the vulnerable A. clanga, and may contribute to the extinction of its populations.