Microsatellite Loci Research Articles

Analysis of 9 microsatellite markers in Kalmyk cattle

A genetic analysis of the Kalmyk cattle population for 9 microsatellite loci: BM1824, BM 2113, INRA023, SPS 115, TGLA 122, TGLA 126, TGLA 227ETH 10, ETH 225 was carried out on the basis of the youth scientific laboratory of molecular genetics of the Kalmyk State University. 60 animals of the Kalmyk breed of breeding reproducer “Plodovitoe” of the Republic of Kalmykia were subjected to genetic testing. The loci INRA 023, TGLA 122, and TGLA 227 turned out to be the most informative for the Kalmyk breed. A comparison of the observed and expected heterozygosity in the Kalmyk breed shows that all 9 microsatellite loci showed an excess of the expected heterozygosity value over the observed one. The number of possible genotypes (Ng) has the highest value at the TGLA 122 locus, the indicator is 171, a smaller number of possible genotypes is observed at the TGLA 126 locus. Thus, microsatellite analysis showed a high level of genetic diversity in the studied herd of Kalmyk cattle. The data obtained indicate the possibility of using microsatellite DNA analysis for genetic examination of breeding animals by origin and analysis of pedigrees in order to study the transmission of traits by inheritance.

Genetic investigation of population structure in Atlantic chub mackerel, Scomber colias Gmelin, 1789 along the West African coast.

Sustainable management of transboundary fish stocks hinges on accurate delineation of population structure. Genetic analysis offers a powerful tool to identify potential subpopulations within a seemingly homogenous stock, facilitating the development of effective, coordinated management strategies across international borders. Along the West African coast, the Atlantic chub mackerel (Scomber colias) is a commercially important and ecologically significant species, yet little is known about its genetic population structure and connectivity. Currently, the stock is managed as a single unit in West African waters despite new research suggesting morphological and adaptive differences. Here, eight microsatellite loci were genotyped on 1,169 individuals distributed across 33 sampling sites from Morocco (27.39°N) to Namibia (22.21°S). Bayesian clustering analysis depicts one homogeneous population across the studied area with null overall differentiation (F ST = 0.0001ns), which suggests panmixia and aligns with the migratory potential of this species. This finding has significant implications for the effective conservation and management of S. colias within a wide scope of its distribution across West African waters from the South of Morocco to the North-Centre of Namibia and underscores the need for increased regional cooperation in fisheries management and conservation.

Microsatellite Analysis of the Genetic Diversity of Mussels from the North-Western Region of the Black Sea

Using four microsatellite DNA markers (<i>Mch 5, Mch 8, MT 203, MT 282</i>), the genetic diversity and population structure of three groups of mussels from the north-western section of the Black Sea were analyzed, and mussels from the Baltic and North Seas were studied as outgroups. By the studied microsatellite (MS) loci, 59 alleles were determined in five mussels' groups (<i>n</i> = 118). The observed heterozygosity (<i>H<sub>O</sub></i>) ranged from 0.083 to 0.538 and was lower than expected (<i>H<sub>E</sub></i> = 0.344-0.858). All mussel groups showed heterozygote deficiency, which suggests the possibility of inbreeding. Weak (<i>F<sub>ST</sub></i> = 0.036-0.048) and moderate (<i>F<sub>ST</sub></i> = 0.054-0.127) genetic differentiation between the studied groups of mussels was revealed. Genetic distances between the mussels' groups from different locations were calculated, which ranged from 0.211 to 1.130. The obtained data were confirmed by cluster and coordinate analyzes carried out using genetic distances between the groups by distribution of alleles frequencies.

Illumina-based transcriptomic analysis of the fast-growing leguminous tree Acacia crassicarpa: functional gene annotation and identification of novel SSR-markers.

Acacia crassicarpa is a fast-growing leguminous tree that is widely cultivated in tropical areas such as Indonesia, Malaysia, Australia, and southern China. This tree has versatile utility in timber, furniture, and pulp production. Illumina sequencing of A. crassicarpa was conducted, and the raw data of 124,410,892 reads were filtered and assembled de novo into 93,317 unigenes, with a total of 84,411,793 bases. Blast2GO annotation, Benchmark Universal Single-Copy Ortholog evaluation, and GO-term classification produced a catalogue of unigenes for studying primary metabolism, phytohormone signaling, and transcription factors. Massive transcriptomic analysis has identified microsatellites composed of simple sequence repeat (SSR) loci representing di-, tri-, and tetranucleotide repeat units in the predicted open reading frames. Polymorphism was induced by PCR amplification of microsatellite loci located in several genes encoding auxin response factors and other transcription factors, which successfully distinguished 16 local trees of A. crassicarpa tested, representing potentially exploitable molecular markers for efficient tree breeding for plantation and biomass exploitation.

ОЦЕНКА ПОДВИДОВОЙ ПРИНАДЛЕЖНОСТИ ПЧЕЛ В ПРИРОДНОМ ПАРКЕ «МУРАДЫМОВСКОЕ УЩЕЛЬЕ»

The article presents the results of a study on the analysis of the subspecies belonging of bee colonies from two apiaries in the Muradymov Gorge Nature Park using nine microsatellite loci of nuclear DNA and the intergenic locus COI-COII of mitochondrial DNA. As a result, it was found that only one family descends on the maternal line from the evolutionary branch M (to which the subspecies dark forest, or Central Russian, bee Apis mellifera mellifera belongs), the remaining families descend from the branch C (includes subspecies A. M. carnica, A. m. ligustica, A. caucasica). The analysis of microsatellite loci of nuclear DNA showed that all families have a hybrid origin or belong to subspecies from the evolutionary branch of C.

Unraveling biotypes of the northern house mosquito, Culex pipiens s.l. (Diptera: Culicidae): molecular differentiation and morphometric analysis.

Geometric morphometrics was used to determine whether geographic isolation could explain differences in wing size and shape between and within continental (27°S to 41°S) and insular (Rapa Nui) populations of Culex pipiens s.s. Linnaeus and their biotypes (f. pipiens and f. molestus). Molecular protocols based on polymorphisms in the second intron of nuclear locus ace-2 (acetylcholinesterase-2) were used to differentiate Cx. pipiens s.s. from Cx. quinquefasciatus Say, and an assay based on polymorphisms in the flanking region of a microsatellite locus (CQ11) was used to identify biotypes. Culex pipiens f. molestus and hybrids shared larval habitats in all continental sites, while Cx. pipiens f. pipiens was found in 5 of the 10 sites. Only biotype molestus was found in Rapa Nui (Easter Island) Pipiens and molestus biotypes occur sympatrically in aboveground locations, and only molestus was found in the underground site (ME). Biotype molestus was dominant in rural locations and preferably anthropophilic. These results agree with the ecological descriptions previously reported for the biotypes of Cx. pipiens s.s. Procrustes ANOVA only showed differences in centroid size between biotypes in females and males and did not show significant differences in wing shape. However, we found significant differences among the geographic areas in the centroid size and wing shape of both females and males. Particularly, the population of Rapa Nui Island had shorter wings than the continental populations. The results highlight the effects of geographic and environmental processes on morphotypes in vector mosquitoes.

First genetic evaluation of a wild population of Crocodylus intermedius: New insights for the recovery of a Critically Endangered species.

During the second third of last century, the Orinoco Crocodile (Crocodylus intermedius) underwent a hunting process driven by the demand from the North American, European, and Japanese leather industry, resulting in a sharp decline of its populations. Currently, only two known remaining populations of this Critically Endangered species persist in the Colombian Orinoquía: in the Guayabero-Duda-Lozada and the Cravo Norte-Ele-Lipa River Systems. The latter has been the only population subject of study, including recent surveys and local conservation initiatives such as egg and hatchling ranching. Despite suggestions for population recovery based on the observed increase in clutches in the area, information regarding its genetic status has been pending assessment. This research aims to provide a genetic characterization of this remaining population and to evaluate the diversity recovered during a period of the egg ranching initiative. For this purpose, we utilized variable molecular markers, specifically 17 microsatellite loci, nuclear DNA. Despite revealing intermediate levels of genetic diversity, we identified an effective population size of 11.5-17, well below the minimum values proposed for short-term subsistence. While no evidence of inbreeding was found, it is acknowledged as a potential risk based on the population's history. Additionally, we detected a historical bottleneck possibly influenced by arid periods affecting the region since the Pleistocene. While the evaluated population presents a unique opportunity for C. intermedius conservation, it also exposes a high risk of entering the extinction vortex. The primary action to be taken is to support the egg and hatchling ranching program, which successfully recovered most of the genetic diversity present in the population.

Spatial prioritisation for crested newt conservation in Luxembourg: Insights from population genetics and species distribution modelling

Abstract The intensification of agriculture and urbanisation has precipitated the decline of numerous species, including the northern crested newt (Triturus cristatus). To mitigate the impacts of habitat degradation and loss, restoration initiatives depend on evidence‐based decision‐making to identify priority sites for conservation efforts. Here, we employed a multidisciplinary approach, drawing on population genetics, species distribution modelling (SDMs) and landscape genetics to develop a spatial prioritisation framework for the creation and/or restoration of crested newt breeding ponds in Luxembourg. For the genetic analyses, more than 1000 crested newts from 98 ponds across Luxembourg were genotyped at 15 microsatellite loci to assess genetic diversity and spatial clustering. The relationship between newt presence and environmental variables was evaluated using SDMs. Priority restoration areas were identified with PRIORITZR, integrating genetic diversity, habitat suitability and connectivity. The population genetic structure was characterised by spatial clustering, which followed an isolation‐by‐distance pattern. Five sites were identified as significantly differentiated from the remaining populations, warranting targeted conservation efforts. Standing water bodies and proximity to the nearest occupied pond were the most important predictors of crested newt presence. The highest gain in habitat suitability through the creation of new ponds was predicted to occur in southwest Luxembourg. Based on the country‐wide predicted occurrence of crested newts and gains in habitat suitability, areas of highest restoration potential were identified in close proximity to existing crested newt populations. The produced maps of priority restoration areas should inform future conservation efforts of the crested newt in Luxembourg. While the importance of preserving genetic diversity is widely recognised, the theory‐practice gap frequently precludes the incorporation of genetic aspects into in situ conservation measures or management policies. This case study demonstrates the integration of multidisciplinary analyses within an evidence‐based framework to guide species conservation efforts.

Оценка уровня гибридизации популяций темной лесной пчелы Apis mellifera mellifera L. в северных районах Башкортостана

In this study the results of analysis of the genetic structure of bees samples from six northern districts of the Republic of Bashkortostan (Baltachevsky, Birsky, Blagoveshchensky, Buraevsky, Tatyshlinsky and Yanaulsky) were presented. First, using the analysis of the intergenic locus COI-COII, belonging to the evolutionary lineages M (A. m. mellifera) or C (A. m. carnica, A. m. caucasica, A. m. ligustica) was established. Then, in bee colonies A. m. mellifera, the level of introgression of the gene pool of evolutionary lineage C was established based on the polymorphism of 9 microsatellite loci of nuclear DNA. As a result, the lowest level of hybridization was detected in samples from Yanaulsky, Tatyshlinsky, Buraevsky and Baltachevsky districts. The highest level of hybridization was detected in colonies from Birsky and Blagoveshchensky districts. In addition, in these two areas, a large number of colonies from the evolutionary lineage C were identified.

Population-genetics analysis of the brown trout broodstock in the “Panjica” hatchery (Serbia) and its conservation applications

Artificial propagation and stocking of brown trout is a standard practice in recreational fishery management. In recent decades, the importance of maintaining intraspecific diversity and protecting locally adapted lineages has been recognized for the species' long-term survival. The first step in selecting donors for stocking involves distinguishing native trout from non-native and introgressed individuals. The established method for discerning Atlantic hatchery strains from the wild populations involves genetic screening of individual diagnostic SNPs and microsatellite assignment tests. This study, using Serbia's Panjica hatchery as an example, illustrates the proper conduct of routine genetic screening for identifying suitable donors for supportive stocking. The broodstock and reference populations were screened using mtDNA control region, LDH nuclear gene, and 12 microsatellite loci to assess the origin, diversity, and inbreeding levels. The analysis revealed only moderate contamination with Atlantic trout and showed the regional origin of the Danubian genes – over 50% of the broodstock was composed of non-introgressed Danubian individuals tracing their origin to the Zapadna Morava River system. Additionally, the study highlighted a considerable discordance between LDH locus and microsatellites in identifying introgressed individuals, raising concerns about the sole reliance on LDH locus for the identification of Atlantic genetic origin in nuclear DNA.

Genetic diversity of a recovering European roller (Coracias garrulus) population from Serbia.

The European Roller (Coracias garrulus), a long-distance migratory bird, faced a considerable decline in breeding pairs throughout Europe at the end of the 20th century. Due to conservation efforts and the installation of nesting boxes, the population of the European Roller in Serbia has made a remarkable recovery. Here, we used the variability of nucleotide sequences of the mitochondrial DNA (mtDNA) control region and 10 microsatellite loci to assess the genetic diversity and structuring, phylogeographic patterns and demographic history of this species using 224 individuals from Serbia. Our results showed moderate level of genetic diversity (HO = 0.392) and a slightly elevated level of inbreeding and homozygosity (FIS = 0.393). Genetic structuring based on microsatellite data indicated three genetic clusters, but without a clear spatial pattern. High haplotype diversity (Hd = 0.987) of the mtDNA control region sequences was detected, and neutrality tests indicated a recent demographic expansion. The phylogeographic analysis, which also included previously published sequences of the mtDNA control region, supported the subdivision into two distinct European and Asian haplogroups (ΦST = 0.712). However, the results of our study showed that a larger number of haplotypes sampled in Serbia are clustered in the Asian haplogroup as compared to previous studies, indicating a historically continuous distribution of this species and possibly a wider distribution of the subspecies Coracias garrulus semenovwi. Our results suggest that the European Roller population in Serbia is genetically stable, with no evidence of recent bottlenecks, and emphasize the importance of artificial nest boxes for promoting and maintaining population dynamics of European Rollers.

Molecular Diversity Analysis of Some Selected Brri Developed Rice Varieties Using Ssr Markers

Molecular characterization of 26 modern rice varieties of Bangladesh was performed using 52 simple sequence repeat markers (SSRs) to estimate the genetic diversity and to reveal genetic relationships among rice cultivars. A total of 156 alleles were detected where the number of alleles per locus generated by each marker varied from 2 to 7, with an average of 3. The band size for a given microsatellite locus ranged from 79 to 278 bp. The polymorphism information content (PIC) for the SSR loci ranged from 0.08 to 0.79, with an average of 0.35. RM566 was the best marker for identification and variability assessment of varieties as revealed by PIC values. A UPGMA dendrogram generated using the NTSYS-pc revealed four clusters with a similarity coefficient of 0.55, whereas phylogenetic cluster analysis of the SSR data based on Nei-genetic distance divided the varieties into three groups. Two- and three-dimensional graphical views of principal coordinate analysis showed the spatial distribution of the varieties and revealed that BRRI dhan56, BRRI dhan51, BRRI dhan52, BRRI dhan61, BRRI dhan62 and BRRI dhan66 were found far away from the centroid of the cluster. The findings of this study are helpful for varietal identification, background selection during backcross breeding and for selecting the suitable genetically diverse parents for crossing programs. SAARC J. Agric., 21(2): 1-14 (2023)

Conservation Genetics of the Loggerhead Sea Turtle, Caretta caretta, from the Central Mediterranean: An Insight into the Species' Reproductive Behaviour in Maltese Waters.

Loggerhead sea turtle, Caretta caretta (Linnaeus, 1758), nestlings were investigated through specimens found dead either after hatching or unhatched (n = 120) from eight nests around the Maltese islands (Central Mediterranean). Molecular genetics was used to conduct maternity and paternity tests of the collected specimens utilizing expanded mitochondrial DNA sequences from the control region (858 bp) and 25 microsatellite loci (12 dinucleotide loci and 13 tetranucleotide loci). Mitochondrial data produced two haplotypes, CC-A2.1 and CC-A3.1, with the most common haplotype being present in seven nests. Microsatellite data revealed the identity of six different females that were involved in the deposition of the eggs in the eight turtle nests analysed. This confirms that two females laid multiple nests. Additionally, microsatellite data allowed for the determination of multiple paternity, with one clutch being sired by two fathers. These results are useful for monitoring the genetic diversity of loggerhead sea turtle nestlings and of the turtle mothers and fathers contributing to future turtle offspring, which rely on Maltese sandy beaches for their successful start to life. Effective conservation management benefits from merging scientific knowledge with effective measures at potential nesting sites to avoid losses of nestlings caused by human negligence.

Microsatellite markers for genotyping of Kodamaea ohmeri: Demonstrating outbreaks based on a multicenter surveillance study in China

Kodamaea ohmeri, an emerging human pathogen, caused both sporadic and nosocomial infections among immunocompromised people with high mortality. However, there is limited research on the molecular epidemiology of K. ohmeri. A total of fifty microsatellite loci were designed based on K. ohmeri type strain NRRL Y-1932 and three loci were finally selected for microsatellite analysis. Non-duplicated K. ohmeri isolates and strains of other species were collected across China as a part of CHIF-NET program for sensitivity and specificity verification. Antifungal susceptibility was determined using Sensititre YeastOne TM YO10. The three loci (P10, P11 and P26), with a cumulative discriminatory power of 0.98, exhibited a prospective specificity and reproducibility in the PCR of 92 K. ohmeri strains from different hospitals. A total of 54 microsatellite types (MT) were identified and most of them distributed sporadically. However, six strains of MT12 clustered in HZ hospital and were isolated in the same department within two months, indicating a potential outbreak. Of seven isolates exhibited MIC values of >8 mg/L for fluconazole, three isolates from LR hospital shared the same genotype of MT44. Herein, we established a set of microsatellite loci for K. ohmeri, as a rapid and specific tool for genotyping K. ohmeri, and identified several potential clusters. This study will help us better understand the molecular epidemiology of the emerging pathogen K. ohmeri.

Genetic footprints of Quaternary climatic fluctuations on the endangered Tuco-tuco-das-dunas (Ctenomys flamarioni)

Abstract The biogeographic history of a species results from responses to past and current processes, and understanding those responses at both temporal scales is essential. We explored phylogeographic and current genetic variation structure in Ctenomys flamarioni—a subterranean rodent that inhabits a small region of the coastal plain of Brazil—and contrasted the roles of “present-day niche suitability” versus “historical climate stability” to understand current and past intraspecific genetic diversity. We used both mitochondrial DNA sequences (concatenated Cytb and D-loop, n = 78) and nuclear DNA (9 microsatellites loci, n = 154) molecular markers for individuals across the entire distribution of the species. We used phylogeographic trees, Bayesian clustering, and haplotype networks to explore genetic variation and diversity. We correlated genetic diversity with measures of current niche suitability and potential areas of Late Quaternary paleostability. Phylogeographic patterns confirmed low levels of genetic diversity structured at different hierarchical levels. We also found a weak but significant pattern of isolation by distance, indicating that geographical distance partially modulates genetic differences among populations. We also observed a positive correlation between genetic diversity and climate paleostability for microsatellites at local and regional scales. However, we found no significant association between genetic diversity and niche suitability for microsatellites at local or regional scales. We observed several signatures of bottleneck and population expansion at different geographic scales. Our results suggest that Quaternary climatic fluctuations—acting together with habitat suitability and other contemporary factors—played a fundamental role in elucidating the evolutionary history of the species—including patterns of historical dispersal, and current patterns of habitat occupation and genetic structure. In addition, based on our molecular and environmental data sets from the across the range of the species and considering the restricted endemism of C. flamarioni, we emphasize the urgency of conservation actions to protect this endangered species.

Microsatellite loci for detecting extrapair offspring in Wood Warbler (Phylloscopus sibilatrix, Aves) nests in the northeast of the species range

Microsatellite loci for detecting extrapair offspring in Wood Warbler (Phylloscopus sibilatrix, Aves) nests in the northeast of the species range

Population genetic patterns across the native and invasive range of a widely distributed seagrass: Phylogeographic structure, invasive history and conservation implications.

The seagrass Zostera japonica is a dramatically declined endemic species in the Northwestern Pacific from the (sub)tropical to temperate areas, however, it is also an introduced species along the Pacific coast of North America from British Columbia to northern California. Understanding the population's genetic patterns can inform the conservation and management of this species. North Pacific. We used sequences of the nuclear rDNA internal transcribed spacer (ITS) and chloroplast trnK intron maturase (matK), and 24 microsatellite loci to survey 34 native and nonnative populations (>1000 individuals) of Z. japonica throughout the entire biogeographic range. We analysed the phylogeographic relationship, population genetic structure and genetic diversity of all populations and inferred possible origins and invasion pathways of the nonnative ones. All markers revealed a surprising and significant deep divergence between northern and southern populations of Z. japonica in the native region separated by a well-established biogeographical boundary. A secondary contact zone was found along the coasts of South Korea and Japan. Nonnative populations were found to originate from the central Pacific coast of Japan with multiple introductions from at least two different source populations, and secondary spread was likely aided by waterfowl. The divergence of the two distinct clades was likely due to the combined effects of historical isolation, adaptation to distinct environments and a contemporary physical barrier created by the Yangtze River, and the warm northward Kuroshio Current led to secondary contact after glacial separation. Existing exchanges among the nonnative populations indicate the potential for persistence and further expansion. This study not only helps to understand the underlying evolutionary potential of a widespread seagrass species following global climate change but also provides valuable insights for conservation and restoration.

Analysis of chickpea (Cicer arietinum L.) genotypes by microsatellite loci of the QTL-hotspot-­region associated with drought tolerance

Purpose. To determine the polymorphism of microsa­tellite loci of the QTL-hotspot-region of linkage group 4, associated with drought tolerance in Ukrainian chickpea varieties. Methods. Extraction and purification of DNA from seedlings using the CTAB method; polymerase chain reaction; horizontal gel electrophoresis; determination of the size of amplification products using the “Image J” prog­ram. Results. Allelic combinations of microsatellite loci ICCM0249, NCPGR127, TAA170, NCPGR21, TA130, STMS11 of the QTL­hotspot­region of linkage group 4 of the chickpea genome were established. It was found that the loci STMS11, NCPGR127, NCPGR21 were not polymorphic within the sample of varieties analyzed, one allele was detected for each locus; two alleles were detected for the loci ICCM0249 and TAA170 and three alleles for the locus TAA130, indica­ting their polymorphism. Conclusions. Microsatellite loci STMS11, NCPGR127, NCPGR21 are non­polymorphic in seven Ukrainian chickpea varieties. Three loci are polymorphic with two alleles for ICCM0249 and TAA170 and three alleles for TAA130. According to the analysis of chickpea varie­ties, five types of allelic combinations of microsatellite loci ICCM0249, NCPGR127, TAA170, NCPGR21, TA130, STMS11 were established. An allele of 185 bp unique to the sample of cultivars studied was identified in the variety ‘Pamiat’

Patterns of evolution in MHC class II DQA and DQB exon 2 genes of Alpine mountain hares, Lepus timidus varronis, and sympatric and parapatric brown hares, L. europaeus, from Switzerland.

In natural populations, hybridization is known to occur between a wide range of species. However, its evolutionary significance is less clear. Genes involved in fighting pathogens are considered excellent candidates for studying adaptive introgression, although both introgression and balancing selection can generate similar patterns of diversity and differentiation. Here, we compared DQA and DQB MHC class II and microsatellite allelic diversity of sympatric and parapatric mountain (Lepus timidus) and brown hare (L. europaeus) populations from Switzerland. We detected higher genetic diversity in brown hares compared to mountain hares at both MHC and microsatellite loci. We consider the observed patterns of microsatellite diversity both for L. europaeus and L. timidus as result of stochastic demographic processes while the pattern of MHC polymorphism of the studied hare populations can be explained by pathogen-driven selection. Rare bidirectional gene flow between both hare species seems to occur specifically for MHC alleles. However, the high number of shared alleles showing similar high frequency in both species suggests that reciprocally exchanged MHC alleles are being maintained via balancing selection. Adaptation to similar pathogen communities can also lead to parallel selection of MHC alleles. Positive selection, recombination and mutations have played different roles in shaping the patterns of MHC allelic diversity in and differentiation between both species. Results for the latter evolutionary forces do not show a better matching between the sympatric populations compared to the parapatric ones, suggesting a minor role of introgression for the observed evolutionary patterns of the studied hare species.

Multilocus phylogeography of Italian Moorish geckos adds insights into the evolutionary history of European populations

AbstractGeckos of clade III of the Tarentola mauritanica species complex are widespread throughout southern Europe and northern Africa. We investigated the genetic variability of the Italian populations by performing a widespread sampling throughout the mainland and the two main islands of Sicily and Sardinia. We analysed 199 newly generated sequences of the mitochondrial 16S rRNA gene and 269 nuclear genotypes inferred from nine microsatellite loci from 307 individuals. We found 13 new mitochondrial haplotypes in Italy, whereas previous findings reported a single haplotype widespread throughout the country and in the rest of Europe, which currently make Italy the centre of genetic diversity of this taxon. There was no evidence of mitochondrial DNA structuring with geographic correlation. At the population genetic level, our multilocus approach based on nuclear markers returned a shallow genetic structure. Nonetheless, we disclosed the presence of at least four distinct genetic clusters (namely the Adriatic, two Tyrrhenian and the Calabrian clusters). Our findings do not support the two hypotheses proposed to explain the low level of mitochondrial polymorphism in this taxon, namely the genetic hitch‐hiking due to selective sweep and the historical human‐mediated colonization hypotheses. Based on the fossil record, the presence in Italy of this taxon since the Pleistocene Epoch is plausible. Given the discordance in genetic structure between mitochondrial and nuclear DNA, the exact role of the Italian Peninsula in shaping the observed patterns of genetic diversity during the Pleistocenic climatic oscillations needs further investigation.