Drainage-independent genetic structure and high genetic diversity of endangered freshwater pearl mussels (<Emphasis Type="Italic">Margaritifera margaritifera</Emphasis>) in northern Europe

Freshwater pearl mussels (Margaritifera margaritifera) are among the most critically threatened bivalve molluscs worldwide. An understanding of spatial patterns of genetic diversity is crucial for the development of integrative conservation strategies. We used microsatellites to study the genetic diversity and differentiation of 14 populations of M. margaritifera in central Sweden, an area which was described as a major secondary contact zone in postglacial colonisation for other species. Genetic diversity of Swedish pearl mussel populations was much greater than in central and southern Europe but similar to the genetic diversity observed in the northeastern portion of their European range. Genetic differentiation among populations was pronounced but to a large extent independent from present-day drainage systems. The complex patterns of genetic diversity and differentiation in pearl mussel seem to be strongly influenced by the species’ high degree of specialisation and extraordinary life history strategy which involves facultative hermaphrodism and an obligatory encystment stage on a host fish. Genetic drift effects and anthropogenic disturbances resulting in reduction of population size and loss of connectivity are less pronounced in northern pearl mussel populations compared to those in central and southern Europe.     

Origin and genetic diversity of mosquitofish (<Emphasis Type="Italic">Gambusia holbrooki</Emphasis>) introduced to Europe

We provide mitochondrial sequence variation of the invasive fish Gambusia holbrooki from 24 European populations, from Portugal to Greece. Phylogeographic structure in Europe was compared with genetic data from native samples (USA) and historical records were reviewed to identify introduction routes. Overall, data agree with records of historical introductions and translocations, and indicate that the most abundant haplotype throughout Europe originated from North Carolina and corresponded to the first introduction in 1921 to Spain, being transferred to Italy in 1922 and to many countries afterwards. Our results also show that at least another independent introduction occurred first in France and subsequently from France to Greece. Haplotypes of G. affinis were not detected in our European sampling effort but historical records and other data suggest that this species was introduced to Italy in 1927 and it might be present. At the continental scale, there is less diversity in Europe than in North America, in agreement with the low number of introduced fish. At the local scale, some European populations gained diversity from multiple introductions and from “de novo” mutations.     

Taxonomy, evolutionary biology and biogeography of South West European Polyommatus coridon (Lepidoptera: Lycaenidae)

Genetic relationships among 15 South West European populations of Polyommatus coridon were evaluated by the electrophoretic study of 16 gene-enzyme systems. Nei's indexes I and D and other statistics demonstrated that two different species are involved, identifiable as Polyommatus coridon (six populations from Southern France and Italy) and Polyommatus caelestissimus (nine populations from Spain), respectively. The Spanish complex is characterized by a different genetic balance, involving much higher degree of interdemic differentiation. It also differs from populations from France and Italy on one alternative locus showing complete fixation. Species-level distinction is independent of haploid chromosome numbers that vary from 87 to 88 in both groups. An evaluation of selective pressures as operated by climatic factors was attempted. The Pleistocene radiation of this group and consequent patterns of the gene flow were reconstructed using spatial autocorrelation analysis.     

Evidence for strong genetic structure in European populations of the little owl Athene noctua

The little owl Athene noctua is a widespread species in Europe. This mainly sedentary owl experienced reduction in population sizes in some areas due to habitat loss and modification of the landscape. To assess the genetic structure of the populations of western and central Europe, we analysed 333 specimens from 15 geographical areas at 13 microsatellite loci. Statistical analyses and Bayesian clustering procedures detected two major genetically distinct clusters, the first distributed from Portugal to the Czech Republic and the second from the Balkans to Italy. The second cluster was further split into three groups, located in Italy, Sardinia and the Balkans. These groups match four previously‐described mtDNA haplogroups, and probably originated from the isolation of little owl populations in Sardinia and in three glacial refugia (Iberia, south Italy and Balkans) during the ice ages. High genetic admixture was recorded in central and northern Europe, probably as a consequence of the expansion from the refugia during interglacial. The main colonization route originated from the Iberian Peninsula towards central and northern Europe. Contact zones with colonization events from Italy and the Balkans were detected respectively in northern Italy and central Europe. Genetic indices show the existence of moderate levels of genetic variability throughout Europe, although evidence of recent evolutionary bottlenecks was found in some populations. Estimation of migration rates and approximate Bayesian computations highlighted the most likely phylogeographical scenario for the current distribution of little owl populations.     

Out of Anatolia: longitudinal gradients in genetic diversity support an eastern origin for a circum-Mediterranean oak gallwasp Andricus quercustozae

Many studies have addressed the latitudinal gradients in intraspecific genetic diversity of European taxa generated during postglacial range expansion from southern refugia. Although Asia Minor is known to be a centre of diversity for many taxa, relatively few studies have considered its potential role as a Pleistocene refugium or a potential source for more ancient westward range expansion into Europe. Here we address these issues for an oak gallwasp, Andricus quercustozae (Hymenoptera: Cynipidae), whose distribution extends from Morocco along the northern coast of the Mediterranean through Turkey to Iran. We use sequence data for a fragment of the mitochondrial gene cytochrome b and allele frequency data for 12 polymorphic allozyme loci to answer the following questions: (1) which regions represent current centres of genetic diversity for A. quercustozae? Do eastern populations represent one refuge or several discrete glacial refugia? (2) Can we infer the timescale and sequence of the colonization processes linking current centres of diversity? Our results suggest that A. quercustozae was present in five distinct refugia (Iberia, Italy, the Balkans, southwestern Turkey and northeastern Turkey) with recent genetic exchange between Italy and Hungary. Genetic diversity is greatest in the Turkish refugia, suggesting that European populations are either (a) derived from Asia Minor, or (b) subject to more frequent population bottlenecks. Although Iberian populations show the lowest diversity for putatively selectively neutral markers, they have colonized a new oak host and represent a genetically and biologically discrete entity within the species.     

Phylogeography of the European brook lamprey (Lampetra planeri) and the European river lamprey (Lampetra fluviatilis) species pair based on mitochondrial data

The European river lamprey Lampetra fluviatilis and the European brook lamprey Lampetra planeri (Block 1784) are classified as a paired species, characterized by notably different life histories but morphological similarities. Previous work has further shown limited genetic differentiation between these two species at the mitochondrial DNA level. Here, we expand on this previous work, which focused on lamprey species from the Iberian Peninsula in the south and mainland Europe in the north, by sequencing three mitochondrial marker regions of Lampetra individuals from five river systems in Ireland and five in southern Italy. Our results corroborate the previously identified pattern of genetic diversity for the species pair. We also show significant genetic differentiation between Irish and mainland European lamprey populations, suggesting another ichthyogeographic district distinct from those previously defined. Finally, our results stress the importance of southern Italian L. planeri populations, which maintain several private alleles and notable genetic diversity.     

Postglacial colonization of Europe by the barbastelle bat: agreement between molecular data and past predictive modelling

The barbastelle (Barbastella barbastellus) is a rare forest bat with a wide distribution in Europe. Here, we combine results from the analysis of two mtDNA fragments with species distribution modelling to determine glacial refugia and postglacial colonization routes. We also investigated whether niche conservatism occurs in this species. Glacial refugia were identified in the three southern European peninsulas: Iberia, Italy and the Balkans. These latter two refugia played a major role in the postglacial colonization process, with their populations expanding to England and central Europe, respectively. Palaeo‐distribution models predicted that suitable climatic conditions existed in the inferred refugia during the last glacial maximum (LGM). Nevertheless, the overlap between the current and the LGM distributions was almost inexistent in Italy and in the Balkans, meaning that B. barbastellus populations were forced to shift range between glacial and interglacial periods, a process that probably caused some local extinctions. In contrast, Iberian populations showed a ‘refugia within refugium’ pattern, with two unconnected areas containing stable populations (populations that subsisted during both glacial and interglacial phases). Moreover, the match between LGM models and the refugial areas determined by molecular analysis supported the hypothesis of niche conservatism in B. barbastellus. We argue that geographic patterns of genetic structuring, altogether with the modelling results, indicate the existence of four management units for conservation: Morocco, Iberia, Italy and UK, and Balkans and central Europe. In addition, all countries sampled possessed unique gene pools, thus stressing the need for the conservation of local populations.     

Genetic diversity and population structure of the endangered insect species Carabus variolosus in its western distribution range: Implications for conservation

Genetic variation was assessed in the endangered species C. variolosus (Coleoptera: Carabidae), which is listed in the European Habitats and Species Directive. Twelve populations from the north-western margin of its range in Germany and France to the border region between Slovenia and Croatia were investigated for variation at 16 allozyme loci. In general, genetic diversity was rather low as indicated by a mean allelic richness of 1.3 alleles per locus, a mean gene diversity (H _E ) of 0.071 and a mean proportion of polymorphic loci (P _95% ) of 16%. Genetic diversity did not change directionally from the margin towards the core of the range. Very high differentiation between populations (overall F _ST  = 0.465), the results of a Mantel test, and poor accordance between geographical and genetic distance suggest a high degree of isolation of the island-like distributed populations. High F _ST values and genetic distance measures, even between geographically close populations within the same drainage, confirm a very low dispersal power of this habitat specialist of headwater areas and swamps in woodlands, even in comparison with other flightless beetles. It is concluded that the majority of populations are demographically independent. Hierarchical F-statistics indicate that almost half of the genetic variance is found between regions and populations. Therefore, we recommend that conservation efforts consider every population as a management unit and aim to maintain as many populations as possible over as many regions as possible.     

Surprising diversity in the Pannonian populations of Marsh Fritillary (Euphydryas aurinia,Lepidoptera: Nymphalidae): Morphometric and molecular aspects

Since genetic variation is the basis of evolutionary potential of a species, its structure needs to be understood. Thus, the aim of this study was to analyze and contrast the structure of genetic and phenotypic variation in the Euphydryas aurinia populations of southeastern central Europe. Genetic variation was studied by two types of molecular genetic markers: mtDNA COI sequences and allozymes. As the great hiatus in the European distribution of E. aurinia is located in the central part of the Carpathian Basin, we expected that the populations East and West to this gap would be highly differentiated. Populations of Central Transdanubia actually represent the easternmost margin of the West European distribution of E. aurinia. In view of the peripheral position of these populations, we supposed to find some genetic sign of local adaptation, as a consequence of diversifying selection and an increased level of fluctuating asymmetry as a result of environmental stress. The analyses of the molecular genetic markers revealed a basic East–West differentiation among the populations of southeastern central Europe which was further structured in the western part of the study area. The results suggested that the genetic differentiation between the two western regions is probably the consequence of diversifying selection. The pattern of phenotypic differentiation among the western populations, however, was different. A geographic cline was revealed (decreasing wing size) toward the eastern margin of the distribution in parallel with increasing fluctuating asymmetry. The conservation inferences of the results are considered.     

Phylogeographic evidence for the postglacial colonization of the North and Baltic Sea coasts from inland glacial refugia by Triglochin maritima L.

We investigated the geographical distribution of genetic variation in 67 individuals of Triglochin maritima from 38 localities across Europe using AFLP markers. Analysis of genetic variation resulted in the recognition of two major genetic groups. Apart from few geographical outliers, these are distributed (1) along the Atlantic coasts of Portugal, Spain and France and (2) in the North Sea area, the Baltic Sea area, at central European inland localities, the northern Adriatic Sea coast and the Mediterranean coast of southwest France. Considering possible range shifts of T. maritima in reaction to Quaternary climatic changes as deduced from the present-day northern temperature limit of the species, Quaternary changes of coastline in the North Sea area and the very recent origin of the Baltic Sea, we conclude that the coastal populations of T. maritima in the North Sea and Baltic Sea areas originated from inland populations.     

Large-scale geographic patterns of genetic variation in Melica nutans, a widespread Eurasian woodland grass

 The geographic distribution of allozyme variation within the Eurasian boreo-nemoreal woodland grass Melica nutans L. has been investigated together with a minor subset of other Melica species. Twenty alleles were found at nine polymorphic loci in M. nutans. Allelic richness was highest in areas central in the species' European distribution, i.e. in southern Fennoscandia. High population densities, reducing the effects of genetic drift, as well as accumulation of variation through long-distance gene-flow from different marginal populations, is proposed to explain high allelic richness in this area. Several alleles showed geographic patterns in distribution and frequency variation. However, these patterns were not congruent, e.g. some alleles appear to have migrated to northern Europe from the south-west whereas others may have spread from the east. Genetic distances between geographic regions, each consisting of 2–6 populations, were generally low between all Fennoscandian, Russian and Siberian regions, but much higher between western and continental European regions. On the population level, cluster analysis grouped populations from Siberia, Russia, coastal and lowland areas in Fennoscandia and British Cumbria into one subcluster whereas other subclusters contained mainly south-west European populations or populations from almost throughout the distribution range. A scenario with several independent glacial refugia in central Europe, south-western Siberia and possibly western Norway, and subsequent colonisation of Fennoscandia mainly from the east, but with some long-distance gene-flow from central Europe, is proposed. Received April 3, 2002; accepted September 17, 2002 Published online: December 11, 2002     

Genetic analyses of the Asian longhorned beetle (Coleoptera,Cerambycidae, <Emphasis Type="Italic">Anoplophora glabripennis</Emphasis>), in North America,Europe and Asia

The Asian longhorned beetle, (Coleoptera, Cerambycidae, Anoplophora glabripennis (Motschulsky)), is endemic to China and Korea and an important invasive insect in North America and Europe. We analyzed mitochondrial DNA sequence data of invasive populations of A. glabripennis in North America and Europe, and microsatellite allele frequency data of beetles from North America. We show that populations in New York City and Long Island NY; New Jersey, Chicago, IL, and Toronto, Canada have limited genetic diversity compared to populations in China. In addition, the data suggest that separate introduction events were responsible for many of the populations in North America and for European populations in Austria, France, Germany and Italy. Populations on Long Island, NY are suspected to have been initiated by the transport of cut wood from New York City. A. glabripennis beetles found in Jersey City, NJ appear to be derived from an expansion of the New York City, NY population, whereas beetles found in Linden, NJ are an expansion from the Carteret, NJ population. Limited genetic diversity did not stop this invasive insect from establishing damaging populations in North America. Founders of introduced A. glabripennis populations in North America and Europe are likely derived from populations in China that are themselves invasive, rendering difficult the identification of source populations. Invasiveness in an insect’s natural range could be an important predictor of potential pest status of introduced populations.     

Microsatellite variation in Bavarian populations of European grayling (Thymallus thymallus): Implications for conservation

European grayling populations in Bavaria have shown steady declines during the last 10–20years. In order to provide guidelines for conservation strategies and future management programs, we investigated the genetic structure of 15 grayling populations originating from three major Central European drainages (the Danube, the Elbe and the Rhine/Main) using 20 microsatellite loci. Genetic divergence between the three drainage systems was substantial as illustrated by highly significant heterogeneity of genotype frequencies, high number of drainage-specific private alleles, high between-drainage F _ST values, high assignment success of individuals to their drainage of origin and the high bootstrap support for the genetic distance based drainage-specific population clusters. In agreement with earlier studies, microsatellites revealed relatively low levels of intrapopulational genetic diversity in comparison to the overall level of variation across populations. Maximum likelihood methods using the coalescent approach revealed that the proportion of common ancestors was generally high in native populations and that the estimates of N _e were correlated with the genetic diversity parameters in all drainages. The number of effective immigrants per generation (N _e m) was less than one for all pairwise comparisons of populations within the drainages, indicating restricted interpopulational gene flow. Based on these findings we recommend a drainage and sub-drainage specific conservation of grayling populations in order to preserve their overall genetic diversity and integrity. For large-scale stocking actions to supplement declining or to restore extinct populations, creation of separate broodstocks for major conservation units (ESUs and MUs) is warranted. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic diversity and ecological differentiation in the endangered fen orchid (<Emphasis Type="Italic">Liparis loeselii</Emphasis>)

Liparis loeselii is a rare and endangered orchid occurring in Europe and north-east America. Genetic diversity and structure of this species in north-west France and the United Kingdom were investigated using amplified fragment length polymorphisms (AFLPs). Although clonality and autogamy are common in L. loeselii, we found moderate to important variability within populations. We observed a significant genetic differentiation between populations occurring in dune slacks and fens. This may be correlated with leaf shape as dune slack individuals are sometimes treated as the distinct variety L. loeselii var. ovata. Genetic differentiation between populations was generally low suggesting that gene flow can occur over long distances and possibly across the English Channel. These results show that populations from dune slacks and fens should be managed separately and that geographically distant populations may be equivalent.     

Landscape genetics structure of European sweet chestnut (<Emphasis Type="Italic">Castanea sativa</Emphasis> Mill): indications for conservation priorities

Sweet chestnut is a tree of great economic (fruit and wood production), ecological, and cultural importance in Europe. A large-scale landscape genetic analysis of natural populations of sweet chestnut across Europe is applied to (1) evaluate the geographic patterns of genetic diversity, (2) identify spatial coincidences between genetic discontinuities and geographic barriers, and (3) propose certain chestnut populations as reservoirs of genetic diversity for conservation and breeding programs. Six polymorphic microsatellite markers were used for genotyping 1608 wild trees sampled in 73 European sites. The Geostatistical IDW technique (ArcGIS 9.3) was used to produce maps of genetic diversity parameters (He, Ar, PAr) and a synthetic map of the population membership (Q value) to the different gene pools. Genetic barriers were investigated using BARRIER 2.2 software and their locations were overlaid on a Digital Elevation Model (GTOPO30). The DIVA-GIS software was used to propose priority areas for conservation. High values of genetic diversity (He) and allelic richness (Ar) were observed in the central area of C. sativa’s European distribution range. The highest values of private allelic richness (PAr) were found in the eastern area. Three main gene pools and a significant genetic barrier separating the eastern from the central and western populations were identified. Areas with high priority for genetic conservation were indicated in Georgia, eastern Turkey, and Italy. Our results increase knowledge of the biogeographic history of C. sativa in Europe, indicate the geographic location of different gene pools, and identify potential priority reservoirs of genetic diversity.     

Phylogeography,population genetics and conservation of the European red deer Cervus elaphus

• 1 During the Last Glacial Maximum, European red deer Cervus elaphus occurred in refugia in Iberia/southern France, Italy, the Balkans and the Carpathians. Most of Europe, including large parts of the east and north‐east, is now inhabited by red deer from the western lineage. The eastern lineage is largely confined to south‐eastern Europe; a third lineage comprises Sardo‐Corsican and Barbary red deer.
• 2 Sardo‐Corsican, Barbary and Mesola red deer are genetically unique units. They exhibit low levels of genetic diversity and deserve particular protection, since conservation strategies should target genetic information.
• 3 Hybridization between sika Cervus nippon and red deer occurs rarely, but may lead to extensive introgression, particularly in parts of the British Isles. Further expansion of both species may lead to increased hybridization in continental Europe.
• 4 Although hunting has an impact on red deer gene pools, the main threat today is habitat fragmentation in human‐dominated landscapes. The resulting increase in genetic drift and inbreeding reduces variability in isolated populations and may lead to inbreeding depression. To support vital meta‐populations, migration corridors should be established.

     

Genetic relationships among some new cat populations sampled in Europe: A spatial autocorrelation analysis

The allelic frequencies of nine Mendelizing genetic characteristics that control coat colour, tabby and length and some skeletal abnormalities have been studied in four feral domestic cat populations, two in the north of Catalonia (Girona and Roses & L’Estartit, northeastern Spain) and two Adriatic Italian populations (Rimini and Venice). Using different genetic and multivariate analyses (Nei’s and Cavalli-Sforza and Edwards’s genetic distances, phenograms and cladograms using different algorithms, strict consensus trees, canonical population, principal coordinates and nonmetric multidimensional scaling analyses), I show the genetic relationships between these populations and other Western European cat populations previously studied. In the Western European area comprising Catalonia, Italy, France and Great Britain, I found significant spatial structure for thet ^b, l andW alleles and for the average correlogram for the seven alleles studied as a whole using a spatial autocorrelation analysis. The genetic distance matrices between these European cat populations also showed a significant correlation with the geographical distance between these populations using Mantel’s test. These analyses showed that in each of these countries, local cat populations have characteristic genetic profiles which were different to neighbouring populations in nearby countries. At least in this area of Western Europe, the geographical distances between cat populations (although the gene flow can be relatively high) is an important factor which can explain differences in allele frequencies between these populations.     

Distinct invasion sources of common ragweed <Emphasis Type="Italic">(Ambrosia artemisiifolia)</Emphasis> in Eastern and Western Europe

The common ragweed (Ambrosia artemisiifolia L.; Asteraceae) is a North American native that is invading Eurasia. Besides its economic impact on crop yield, it presents a major health problem because of its highly allergenic pollen. The plant was imported inadvertently to Europe in the eighteenth century and has become invasive in several countries. By analyzing French and North American populations, it was previously shown that French populations were best described as a mixture of native sources and that range expansion in France probably involved sequential bottlenecks. Here, our aim was to determine whether Eastern European populations of A. artemisiifolia originated from the previously established French populations or from independent trans-Atlantic colonization events. We used nuclear microsatellite markers to elucidate the relationships among populations from Eastern and Western Europe in relation to populations from a broad survey across the native North American range. We found that A. artemisiifolia from Eastern Europe did not originate from the earlier established French populations but rather represents multiple independent introductions from other sources, or introductions from a not yet identified highly diverse native population. Eastern European populations show comparable amounts of genetic variability as do previously characterized French and North American populations, but analyses of population structure clearly distinguish the two European groups. This suggests separate introductions in Eastern and Western Europe as well as divergent sources for these two invasions, possibly as a result of distinct rules for trade and exchange for Eastern Europe during most of the twentieth century.     

High levels of genetic diversity in marginal populations of the marsh orchid Dactylorhiza majalis subsp. majalis

Because of harsh conditions, suboptimal habitat quality and poor connectivity to other populations, plant populations at the margin of a distribution are expected to be less genetically diverse, but to be more divergent from each other than populations in the centre of a distribution. In northern Europe, northern marginal populations may also be younger than populations further to the south, and may have had less time to accumulate genetic diversity by mutation and gene flow. However, orchids have very small seeds, which are easily dispersed long distances by wind, and orchids are therefore expected to show less differentiation between marginal and central populations than other groups of seed plants. Here, we analysed whether Scandinavian populations of the tetraploid marsh orchid Dactylorhiza majalis subsp. majalis differ from central European populations in genetic diversity patterns. A total of 220 plants from eight central European and ten Scandinavian populations was examined for variation at five nuclear microsatellite loci, nuclear ITS and 13 polymorphic sites in noncoding regions of the plastid genome. The total genetic diversity was slightly lower in Scandinavia than in central Europe, both in plastid and nuclear markers, but the differences were small. Also, the Scandinavian populations were less diverse and somewhat more strongly differentiated from each other than the central European ones. Dactylorhiza majalis subsp. majalis has apparently colonized Scandinavia on multiple independent occasions and from different source areas in the south. Seed flow between Scandinavian populations has still not fully erased the patterns imprinted by early colonization. Our results suggest that marginal populations of orchids may be as important as central ones in preserving genetic diversity through Pleistocene glacial cycles. We also predict that orchids with their light seeds are better adapted than many other plants to respond to future climate changes by dispersing into new suitable areas.     

Global diversity and genetic contributions of chicken populations from African,Asian and European regions

Genetic diversity and population structure of 113 chicken populations from Africa, Asia and Europe were studied using 29 microsatellite markers. Among these, three populations of wild chickens and nine commercial purebreds were used as reference populations for comparison. Compared to commercial lines and chickens sampled from the European region, high mean numbers of alleles and a high degree of heterozygosity were found in Asian and African chickens as well as in Red Junglefowl. Population differentiation (F_ST) was higher among European breeds and commercial lines than among African, Asian and Red Junglefowl populations. Neighbour‐Net genetic clustering and structure analysis revealed two main groups of Asian and north‐west European breeds, whereas African populations overlap with other breeds from Eastern Europe and the Mediterranean region. Broilers and brown egg layers were situated between the Asian and north‐west European clusters. structure analysis confirmed a lower degree of population stratification in African and Asian chickens than in European breeds. High genetic differentiation and low genetic contributions to global diversity have been observed for single European breeds. Populations with low genetic variability have also shown a low genetic contribution to a core set of diversity in attaining maximum genetic variation present from the total populations. This may indicate that conservation measures in Europe should pay special attention to preserving as many single chicken breeds as possible to maintain maximum genetic diversity given that higher genetic variations come from differentiation between breeds.