Applicability of Rabbit Microsatellite Primers for Studies of Hybridisation between an Introduced and a Native Hare Species

Abstract

Introduced species may hybridise with relatives in the native fauna or flora and thereby compete for matings and transmit alien DNA. Such interference may contaminate unique genepools, disturb existing ecological balances and may ultimately result in the extinction of the native species. In Sweden, the introduced brown hare (Lepus europaeus Pall.) hybridise with the native mountain hare (L. timidus L.), both relatively common members of the present Swedish fauna. This hybridisation has resulted in the transmission of mitochondrial DNA from the mountain hare to the brown hare, but absence of species differences in karyotype and allozymes have prevented investigations of the amount of nuclear gene flow. More polymorphic genetic markers are needed to analyse evidence of hybridisation in the nuclear genome. The conservation of microsatellite loci across taxa usually enables PCR amplification of microsatellites in closely related species with the same primers. We have used five microsatellite primer pairs, developed for the European wild rabbit (Oryctolagus cuniculus L.) to amplify microsatellites in the two species of hare in Sweden. The obtained allelic variation was used to construct a genetic distance tree based on the amount of shared alleles between all pairs of individuals (shared‐allele index). This method offered sufficient differences to arrange all individuals in two groups, one for each species. Identification of individual hybrids based on the number of alleles shared between the species is not possible with these five microsatellite markers.

On shortcomings of using mtDNA sequence divergence for the systematics of hares (genus Lepus): An example from cape hares

“…First results from microsatellites, however, do not look very convincing. While Andersson et al (1999) did not find diagnostic microsatellite loci for distinguishing L. europaeus and L. timidus from Sweden, Estonba et al (2006) claimed to have shown a distinction among three Iberian hare species (i.e., L. europaeus, Lepus granatensis, Lepus castroviejoi) with six microsatellite loci. But their cluster (NJ) analysis (Fig.…”

Molecular Approaches Revealing Prehistoric, Historic, or Recent Translocations and Introductions of Hares (Genus Lepus) by Humans

“…Highly variable nuclear DNA markers, such as microsatellite loci, would be desirable for cross checking particularly in regions that are under suspicion of harboring mtDNA introgression among hares, but are currently not elaborated. A microsatellite approach based on five loci did not succeed in unambiguously identifying hybrids of brown and mountain hares (L. europaeus and L. timidus) in southern Scandinavia (see [4]). Application of more loci might produce alleles that are diagnostic for several hare species, or might, in combination with appropriate statistics, enable population assignment.…”

Landscape fragmentation and habitat suitability in endangered Italian hare (Lepus corsicanus) and European hare (Lepus europaeus) populations

“…Three of these (Sat5, Sat12, Sat13) were isolated in rabbit (Mougel et al 1997) but gave good results in hare, too (Andersson et al 1999;Thulin et al 2006). Remaining five microsatellite loci (Lsa1, Lsa2, Lsa3, Lsa6, Lsa8) were identified in L. capensis by Kryger et al (2002) and used in several hare population genetic studies (Thulin et al 2006).…”