Microsatellite analysis of population structure and genetic differentiation within and between populations of the root vole,Microtus oeconomusin the Netherlands

Abstract

Eight microsatellite markers for the root vole (Microtus oeconomus) were developed to assess the amount of genetic variation for nine Dutch root vole populations from four different regions, and to evaluate the degree of differentiation and isolation. All eight microsatellite loci were found to be highly variable with observed heterozygosity values ranging from 0.61 to 0.82. These values are similar to those observed for more distant populations from Norway, Finland and Germany. Therefore, the populations seem not particularly depauperate of genetic variation at the microsatellite level. Genetically, the Dutch populations were found to have diverged considerably. Pairwise comparisons of all populations studied revealed FST values significantly greater than zero for most comparisons. However, the magnitude of these values considerably depends on the compared population pair. The level of differentiation between local populations within Dutch regions is generally significantly lower than the differentiation between Dutch regions. The level of differentiation between Dutch regions, however, is not significantly different from that between populations of larger geographical distance. This implies that the regional Dutch populations are both isolated from each other and from other European populations. The observation that even local populations show low but significant genetic differentiation may be indicative for progressive isolation of these populations.

Genetic structure, habitat fragmentation and bottlenecks in Danish bank voles (Clethrionomys glareolus)

“…The high level of genetic diversity, measured as the number of alleles per locus and the heterozygosity, was similar to levels reported for microsatellites in the same species (H e =0.76) in southern Germany and Switzerland and similar to levels of genetic diversity found in other species of small rodents in Europe ( Van de Zande et al 2000;Ishibashi et al 1999;Harr et al 2000;Dallas et al 1995).…”

“…Populations of wild house mice (Mus musculus and Mus domesticus) in Denmark (Dallas et al 1995) and a Robertsonian race of house mice (Mus domesticus) (Dallas et al 1998) were shown to be highly subdivided genetically based on microsatellite markers. In the root vole Microtus oeconomus microsatellite analysis has shown that nine Dutch populations have diverged considerably and were isolated from each other as well as other European populations (Van de Zande et al 2000). Ehrich et al (2001) demonstrated a clear genetic differentiation among geographical regions but a weaker differentiation between localities within regions of the lemming (Dicrostonyx groenlandicus).…”

Site occupancy response to natural and anthropogenic disturbances of root vole: Conservation problem of a vulnerable relict subspecies

“…Natural and human disturbances may reduce the availability of the species' suitable habitats, therefore the lower local density and the shrinkage of genetic variability increase the risk of local extinction. Previous studies concerning with the population genetic structure of the species have already drawn the attention to this problem (Czajkowska et al 2010;Leijs et al 1999;Papp et al 2000;Van de Zande et al 2000).…”

Single-tube HotSHOT technique for the collection, preservation and PCR-ready DNA preparation of faecal samples: the threatened Cabrera's vole as a model

“…Possible contamination was monitored by using a number of extraction blanks at all extraction steps. PCR amplification, sequencing and genotyping Each 30 μL PCR mixture consisted of 2 μL of gDNA (from tissue or faecal samples), together with the PCR mixture containing primer pairs (1) MiKa1 (5′-ATTACTCCTT-TAAACCATGG-3′) and MiKa2 (5′-CTAATAGACAAAA-TAGGGATGGGG-3′) (Alasaad et al 2011) for mitochondrial control region DNA amplification; or (2) Moe1F (5′-TGGTTGTTCTGTGGTGAATACAG-3′; labelled with the fluorescent 6-FAM) and Moe1R (5′-ACAGTAAGCAGTTTATCCACAAACC-3′) (Van De Zande et al 2000) for Moe1 microsatellite locus amplification (0.25 μM of each primer), 0.12 mM dNTP, 3 μL of 1× kit-supplied PCR buffer, 1.5 mM MgCl 2 , 0.4% BSA, 1.5 μL DMSO and 0.2 μL (0.2 U/reaction) Taq polymerase (Bioline). Samples were subjected to the following thermal profile for amplification in a 2720 thermal cycler PTC-0200 DNA Engine thermal cycler (Bio-Rad): 4 min at 94°C (initial denaturation), followed by 40 cycles of three steps of 1 min at 94°C (denaturation), 1 min at 55°C (annealing for Mika1 and MiKa2 primers) or at 60°C (annealing for Moe1F and Moe1R primers), and 50 s at 72°C (extension), before a final elongation of 5 min at 72°C.…”