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Neogene orogenesis and climatic cycles strongly influenced inter- and intraspecific differentiation and variability of taxa. In this study, we focused on the southern margin of the western Palaearctic, known to be a geographically complex region. We performed mitochondrial DNA analyses of Buthus scorpions from the Moroccan Atlas Mountains, from the Hoggar Mountains in Algeria and from Tunisia, Iberia and Israel. Molecular species delimitation suggests the existence of ≥ 24 molecular operational taxonomic units. The data confirm complex differentiation patterns across the Atlas Mountains of Morocco, but structures in Iberia, Algeria and Tunisia have considerably lower complexity. This identifies the Atlas Mountain region as the most important differentiation centre of Buthus scorpions. Samples from the Hoggar Mountains (southern Algeria) cluster with those from the southernmost parts of Morocco in the middle and upper parts of the Draa Valley. This reinforces a recent connection of these regions. Samples from Israel are genetically similar to individuals from eastern Algeria and Tunisia. This suggests a widespread group across major parts of North Africa. Divergence time estimates indicate that differentiation in the genus began during the late Miocene, a period characterized by strong tectonic activities in this region. Further differentiation could be linked to subsequent climatic changes that have occurred since the end of the Miocene, with an increasing aridification of the Moroccan area. This also produced many microrefugia in the mountains of the area during the Pleistocene climatic fluctuations.

The geographical distributions of species associated with European temperate broadleaf forests have been significantly influenced by glacial–interglacial cycles. During glacial periods, these species persisted in Mediterranean and extra-Mediterranean refugia and later, during interglacial periods, expanded northwards. The widespread saproxylic beetle Bolitophagus reticulatus depends closely on European temperate broadleaf forests. It usually develops in the tinder fungus Fomes fomentarius, a major decomposer of broadleaf-wood. We sampled B. reticulatus in sporocarps from European beech (Fagus sylvatica) and Oriental beech (Fagus orientalis) across Europe and the Caucasus region. We analysed mitochondrial gene sequences (cox1, cox2, cob) and 17 microsatellites to reconstruct the geographical distribution of glacial refugia and postglacial recolonization pathways. We found only marginal genetic differentiation of B. reticulatus, except for a significant split between populations of the Caucasus region and Europe. This indicates the existence of past refugia south of the Great Caucasus, and a contact zone with European populations in the Crimean region. Further potential refugia might have been located at the foothills of the Pyrenees and in the Balkan region. Our genetic data suggest a phalanx-wise recolonization of Europe, a reflection of the high mobility of B. reticulatus.

Species demanding specific habitat requirements suffer, particularly under environmental changes. The smallest owl of Africa, the Sokoke Scops Owl (Otus ireneae), occurs exclusively in East African coastal forests. To understand the movement behaviour and habitat demands of O. ireneae, we combined data from radio-tracking and remote sensing to calculate Species Distribution Models across the Arabuko Sokoke forest in southern Kenya. Based on these data, we estimated the local population size and projected the distribution of current suitable habitats. We found that the species occurs only in Cynometra woodland with large old trees and dense vegetation. Based on home range sizes and the distribution of suitable forest habitats, the local population size was estimated at O. ireneae, and of other specialist forest species, even further. Due to their close connection with intact Cynometra forest, O. ireneae is an excellent indicator of intact forest remnants. In addition, this species is a suitable flagship for the promotion and conservation of the last remaining coastal forests of East Africa.

Different methods to measure species behaviour and space use may produce diverging results, and provide advantages and shortcomings. Data from spot-mapping of animals might be affected from restricted detectability of individuals in dense vegetation, while radio-tracking provides a less biased (or even unbiased) measure of space use. Here we compare results from spot-mapping and telemetry of four family groups (i.e. five individuals belonging to these four family groups), respectively of the Kenyan endemic cooperative-breeding bird Hinde's babbler, Turdoides hindei. Data from spot-mapping showed that the space use of T. hindei is mostly restricted to riparian vegetation. Home-range sizes calculated from telemetry were five times larger if compared with data obtained from spot-mapping. Telemetry data showed that T. hindei also moves across agricultural land, and mean and maximum displacements are larger if compared with data obtained from spot-mapping. Several reasons might lead to these differences: (1) Telemetry data also consider rare long-distance excursions, while (2) observers of spot-mapping might fail to observe long-distance movements and thus underestimate home-range sizes as well as displacement distances; (3) results from telemetry might become blurred from measurement error during the triangulation of fixes. Our study confirms that both methods provide advantages, but also shortcomings, which need to be considered when selecting a method to elaborate a research question.

International audience; Pleistocene glaciations had significant effects on the distribution and evolution of species inhabiting the Holarctic region. Phylogeographic studies concerning the entire region are still rare. Here, we compared global phylogeographic patterns of one boreo-montane and one boreo-temperate butterflies with largely overlapping distribution ranges across the Northern Hemisphere, but with different levels of range fragmentation and food specialization. We reconstructed the global phylogeographic history of the boreo-montane specialist Boloria eunomia (n = 223) and of the boreo-temperate generalist Boloria selene (n = 106) based on mitochondrial and nuclear DNA markers, and with species distribution modelling (SDM). According to the genetic structures obtained, both species show a Siberian origin and considerable split among populations from Nearctic and Palaearctic regions. According to SDMs and molecular data, both butterflies could inhabit vast areas during the moderate glacials. In the case of B. selene, high haplotype diversity and low geographic structure suggest long-lasting interconnected gene flow among populations. A stronger geographic struc-turing between populations was identified in the specialist B. eunomia, presumably due to the less widespread, heterogeneously distributed food resources, associated with cooler and more humid climatic conditions. Populations of both species show opposite patterns across major parts of North America and in the case of B. eunomia also across Asia. Our data underline the relevance to cover entire distribution ranges to reconstruct the correct phylogeographic history of species.

Central European calcareous grasslands are particularly rich in species, harbouring an endangered flora and fauna. In this study, we analyse how invertebrates, like ground beetles, react on vegetation changes in these habitats. We ask if and how species of both groups may react on local environmental changes associated with conservation management regimes. For this reason, vascular plant and ground beetle communities were assessed at six semi-natural calcareous grasslands in Luxembourg before (1988–1992) and after (2008–2010) restoration had started. All studied grassland sites are protected as nature reserves and managed to preserve an open and diverse plant species composition. Our data indicate diverging trends for plant and ground beetle composition: Richness and abundance of species increased for plants and decreased for ground beetles, particularly for forest and Red List species. Functional diversity of plants and ground beetles remained stable, although the abundance of nitrogen-loving plant species increased. In ground beetles, we identified increased proportions of species with strong habitat associations. These findings underline that plant species, in particular typical calcareous grassland species, benefited from ongoing conservation management, while many ground beetle species, mostly the non-grassland specialists, suffered under the applied management regimes. These results highlight the general dilemma in nature conservation of how to prioritise management regimes in situ.

Tinamous (Tinamidae) represent one of the most ancient living avian lineages but their life history traits are relatively unstudied. Here we identified microsatellite loci for two sympatric tinamou species, the Ornate Tinamou ( Nothoprocta ornata) and the Darwin's Nothura ( Nothura darwinii) from low coverage Illumina sequencing of genomic DNA. The experiment yielded a large number of candidate loci. We designed primers and tested them for successful amplification in 1 to 2 populations of the target species, tested for deviation from the Hardy–Weinberg equilibrium and the presence of null alleles, the levels of polymorphism and potential cross-amplification. All 30 and 24 loci amplified consistently, in the Ornate Tinamou and in Darwin's Nothura, respectively. In the Ornate Tinamou, 25 loci were polymorphic and in the Darwin's Nothura 12, with 2 to 14 alleles per locus in both species. The expected heterozygosity ranged from 0.054 to 0.917 in the Ornate Tinamou and from 0.044 to 0.908 in the Darwin's Nothura. 23 (40%) of 54 loci were successfully cross-amplified. These newly discovered, polymorphic microsatellite loci represent a valuable tool for future studies on social behaviour, parentage and genetic population structure in tinamous.

AimIdentification of potential glacial refugia and post-glacial colonization processes of a flightless, cold-adapted ground beetleLocationCentral and eastern Europe.MethodsWe analysed the genetic structure of 33 Carabus sylvestris populations sampled across its entire distribution range using nuclear and mitochondrial markers. We further compiled occurrence records to develop species distribution models to predict distribution ranges for the last glacial period and the present based on the species’ current climatic niche.ResultsDistinct genetic lineages were detected for a number of mountain ranges and were congruent for both molecular marker systems. Most genetic splits were the results of vicariance, whereas dispersal was rare. Our models suggest that the species’ distribution range was larger and more interconnected in the past.Main conclusionsOur data support multiple glacial refugia for C. sylvestris, some of which were located north of the Alps. Some lower mountain ranges were likely recolonized post-glacially.

The distributions of European high mountain species are often characterised by small and geographically isolated populations and, in many cases, have highly complex biogeographic histories. The butterfly genus Erebia represents one of the best examples for small-scale diversification in the European high mountain systems and therefore to understand speciation processes and associated range dynamics of high mountain species. In this study, we analysed 17 polymorphic allozyme loci of 1731 individuals from 49 populations representing four species, one of which has three subspecies: Erebia nivalis; Erebia tyndarus; Erebia ottomana; and Erebia cassioides cassioides, Erebia cassioides arvernensis, and Erebia cassioides neleus. Samples were collected in the high mountain systems of Europe (i.e. Pyrenees, Massif Central, Alps, Apennines, Carpathians, Balkan high mountains). Genetic analyses supported all previously accepted species. However, the genetic differentiation within E. cassioides sensu lato into three geographically delimited groups is justifying species rank: E. arvernensis distributed in the Pyrenees, Massif Central and western Alps; E. cassioides sensu stricto in the eastern Alps and Apennines; and E. neleus in the Balkan mountains and the south-western Carpathians. While the differentiation between western Alps and Massif Central as well as eastern Alps and Apennines was low, the Pyrenees as well as the south-western Carpathians were significantly differentiated from the other regions within the respective taxon. In general, the differentiation among the populations of E. neleus was stronger than between populations of the other taxa. Within E. cassioides, we found a west-east gradient of genetic similarity over the eastern Alps. Based on the obtained genetic structures, we are able to delineate glacial refugia and interglacial range modifications. Based on the genetic structures and genetic diversity patterns, we conclude that, triggered by the glacial-interglacial cycles, repeated range modifications have taken place with subsequent differentiation and speciation in the region of the Alps and Balkans. Colonisations to Pyrenees (E. arvernensis pseudomurina, E. arvernensis pseudocarmenta), Massif Central (E. ottomana tardenota, E. a. arvernensis) and Apennines (E. cassioides majellana) appear to be recent and most probably not older than the last interglacial period.

Conflicts between human needs and nature conservation are exceptionally pronounced along rivers in tropical Kenya, where riparian ecosystems create important retreats for many endangered species, but also provide important ecosystem services for the local human population. This situation has led to a landscape mosaic consisting of dense thickets and agricultural plots. We assessed the occurrence of the Kenyan endemic riparian bird species HindesBabbler Turdoides hindei along three rivers in south-east Kenya. We analysed the landscape coverage within habitat circles of 220 m radius, which are occupied and unoccupied (the latter randomly selected along the rivers) by our targeted bird species. Based on these data we calculated habitat preferences and population structure of T. hindei. Our data reveal that its occurrence probability increased with cov- erage of thickets. Furthermore, geographic distances among local populations of T. hindei decreased with thicket coverage and vice versa. These data reveal the relevance of thicket coverage as a key factor for the occurrence of T. hindei, influencing its population structure. However, most of the thicket patches mapped along the three rivers are small and geographically isolated from each other. Further deforestation might lead to additional reduction of the population size and abundance of T. hindei, and may ultimately lead to local extinction of this, and other endangered species adapted on riparian thickets. This

Many sedentary species with specific habitat demands occur in isolated populations and thus show strong intraspecific genetic differentiation, shaped either by geographical isolation or by local ecological adaptation to specific environmental conditions. The water-lily leaf beetle Galerucella nymphaeae has two ecotypes: one only found feeding as a specialist on just a few host plant species of the family Polygonaceae, and the other on various representatives of the family Nymphaeaceae as a generalist. To test whether the differentiation in ecotypes is also reflected in the population genetic structure, we combined amplified fragment length polymorhism data (> 500 polymorphic fragments; four sampling sites, 27–31 individuals each) with ex-situ feeding experiments (ten individuals from each site). We found that both ecotypes are genetically well differentiated. Furthermore, the genetic divergence between the two studied generalist Nymphaeaceae ecotype populations was also high. In contrast, the two specialist Polygonaceae ecotype populations clustered closely together (thus not depending on geographical isolation), indicating an ecology-driven genetic structuring. Our ex-situ reciprocal feeding experiment revealed that only individuals of the Polygonaceae ecotype are strictly dependent on their host plant family: individuals from populations feeding Polygonaceae are less likely to feed on species from the family Nymphaeaceae than the reverse. Our data show that host preference may drive genetic differentiation even more strongly than simple geographical distance.

Thickets along rivers in Eastern Kenya are important habitats for many endangered species. These habitats also provide fundamental ecosystem services for humans. Intense anthropogenic activities during the past decades have caused a severe reduction of this vegetation and resulted in fragmentation of the remaining thicket patches. We assessed the occurrence of the Kenyan endemic bird species Hinde's Babbler (Turdoides hindei) in a highly fragmented environment and performed detailed land use mapping along the Nzeeu River in East Kenya. We measured the time the birds spent in thicket patches, which differed in their habitat setting: pristine versus surrogate vegetation, different habitat size and different edge-size-ratio. Further, we identified areas of potential conflicts between human activities and our target species. Four T. hindei family groups were observed, mostly in invasive Lantana camara patches. Habitat size and edge-size-ratio of the respective thicket patches revealed a significant impact on the duration of stay of T. hindei with disproportional longer stays in small habitat patches and in patches with larger edge-size-ratio than in rather large patches or thickets with small edge-size-ratio. The 75%- and 95%-kernels showed no overlap between family groups and only marginal overlap with the 75%-kernels of human disturbances. Our data show that the invasive L. camara thickets (even small patches with high edge-size ratio) are a suitable surrogate habitat for the Kenyan endemic for T. hindei. The birds avoid open land likely because of higher predation pressure outside of thickets. Limited overlap between zones of human activity and the occurrence of T. hindei may be a response either to lacking thickets in these areas, and/or an adaptation to elevated hunting pressure in these zones. Therefore, the transformation of thickets into open agricultural land has a negative impact on the persistence of T. hindei. For the preservation of the remaining T. hindei family groups in our study area we suggest to establish an interconnected network of thicket patches, as the high mobility of the species allows persistence in such patchy environments.

Anthropogenic activities caused a severe loss of pristine habitats alongside with fragmentation of remaining habitats and the deterioration of habitat quality. The Arabuko Sokoke forest represents the largest remnant of East African coastal forest. Despite conservation efforts to maintain biodiversity in this forest, populations of several species are declining, including flagship species like the endemic Sokoke Scops Owl Otus ireneae. In this study, we assessed the presence of O. ireneae based on playback technique to identify the occurrence of this species across the forest. We then analysed habitat parameters at sites occupied and non-occupied by the owl species. Our data show that O. ireneae occurs restricted to one single forest type, the Cynometra woodland. Results obtained from parametric one-way ANOVA and structural equation modelling reveal that large old Cynometra webberi trees and dead wood are key requirements for the occurrence of this owl species. However, large C. webberi trees are currently illegally logged by the forest adjacent human population to produce timber, carvings, poles. Charcoal and dead wood represents a major energy source for households. Otus ireneae, being the smallest owl of East Africa may serve as a charismatic flag ship species to promote conservation of this endangered coastal forest, and furthermore as umbrella to conserve other forest species with similar habitat requirements.

East-African mountain forest species often occur in small and isolated populations, whereas species inhabiting the dry lowland savannahs exist in large and interconnected population networks. Taxa with closely related highland and lowland species, such as the East- African White-eye birds, allow testing for the potential effects of the two contrasting distribution patterns, moun- tain disjunction versus lowland panmixia. In this study, we compare the population genetic and bioacoustic differen- tiation of two representatives of the genus Zosterops: Zo- sterops poliogaster is exclusively found in forests at higher elevations; in comparison, Zosterops abyssinicus, only occurs in the dry and warm lowland savannahs. Both species were analysed across a similar geographical scale. Population genetic differentiation was inferred using the same set of 15 polymorphic microsatellite loci for both species. In addition, we quantitatively analyzed bioacoustic traits. Both data sets indicate a strong population differ- entiation among populations of the highland species, but an absence of differentiation in the lowland species. In addi- tion, the lowland Z. abyssinicus was characterised by a twofold higher genetic diversity than detected for the highland Z. poliogaster. These two contrasting intraspecific population structures may reflect the opposite ecology and distribution of these species: the strong population isolation of Z. poliogaster resulting from long-term restriction to the cool and moist mountain forests at higher elevations has led to strong differentiation among local populations and resulted in a comparatively low level of intraspecific var- iability. In contrast, population panmixia in the lowland Z. abyssinicus provides a high level of gene flow allowing the maintenance of high genetic diversity and avoiding strong population structuring. These findings need to be consid- ered when planning conservation actions.

Conservation International's biodiversity hotspots are areas of high vascular plant endemism combined with high levels of habitat destruction and land use change. Although such hotspots have also been shown to be centres for terrestrial vertebrate endemism, much less is known about how well these areas function as hotspots for other less well-studied groups, including the hyperdiverse arthropods, other invertebrates and fungi. Because there is a close evolutionary and ecological relationship between insects and plants, we suggest that the potential role of plants as umbrella species for herbivorous insects, potentially herbivorous fungi and nematodes, and parasitic insects should be explored. Finally, we reflect on the increasing social, economic, human conflict and governance issues and the impacts of increasing land use change and global climate change that threaten the biodiversity hotspot system.

The Eastern Afromontane biodiversity hotspot consists of isolated mountain massifs embedded within the dry lowland savannas of East Africa and of which the peaks and ridges are covered by cloud forest remnants. These cloud forests are home to the Mountain White-eye (Zosterops poliogaster), while three congeneric species (Abyssinian White-eye, Zosterops abyssinicus; Yellow White-eye, Zosterops senegalensis; Pemba White-eye, Zosterops vaughani) inhabit the adjacent lowland savannas. We sampled individuals of all four species across Kenya to analyse interspecific genetic relationships as well as intraspecific differentiation among mountain populations of Z. poliogaster. While the level of genetic differentiation among the four species was rather low, genetic differentiation within Z. poliogaster was very high, even between geographically neighbouring populations. Overall, levels of genetic variation varied strongly across all four species, with much higher diversity detected within the three lowland ones. The highland species was characterised by numerous private alleles that were geographically restricted at populations from single mountains, some of which showed evidence of recent population bottlenecks. We conclude that Z. poliogaster populations are both of high conservation value and conservation concern, given the high proportion of endemic alleles and the genetic signatures of high genetic drift and low gene flow that are typical for small and isolated populations.

Aim Thermophilic species persisted in southern refugia during the cold phases of the Pleistocene, and expanded northwards during warming. These processes caused genetic imprints, such as a differentiation of genetic lineages and a loss of genetic diversity in the wake of (re)colonization. We used molecular markers and species distribution models (SDMs) to study the impact of range dynamics on the common wall lizard, Podarcis muralis, from southern refugia to the northern range margin. Location Parts of the Western Palaearctic. Methods We genotyped 10 polymorphic microsatellites in 282 individuals of P. muralis and sequenced the mitochondrial DNA (mtDNA) cytochrome b gene to study the genetic structure, divergence times and ancestral distributions. Furthermore, we generated SDMs for climate scenarios for 6 and 21 ka derived from two different global circulation models. Results We detected two major mtDNA lineages – a western France clade (Pyrenees to Brittany), and an eastern France clade (southern France to Germany, Belgium and Luxembourg). This split was dated to c. 1.23 Ma. The latter clade was divided into two subclades, which diverged c. 0.38 Ma. Genetic diversity of microsatellites within each clade was nested and showed a significant loss of genetic diversity from south to north, a strong pattern of allele surfing across nearly all loci, and an increase in genetic differentiation towards the northern range margin. Results from SDMs suggest that southward range retraction during the late glacial period split the distribution into geographically distinct refugia. Main conclusions The strong genetic differentiation mirrors the effects of long-term isolation of P. muralis in multiple refugia. Post-glacial recolonization of Northern Europe has taken place from two distinct refugia, most probably along river systems (Rh^ Rhine, Moselle) and along the Atlantic coastline, with subsequent nested elimination of genetic diversity and increasing genetic differentiation at the northern range margin.

Tinamous have male-only parental care. For their mating system, simultaneous polygyny and sequential polyandry is proposed as an optimal model and is confirmed in all species according to the existing literature, except in the Ornate Tinamou. For the Ornate Tinamou a monogamous mating system is reported. However, this knowledge is based only on few direct observations in the field. Social monogamy in association with promiscuity would probably expose males to cuckoldry and, at the same time, constrain additional mating opportunities for them. To understand the evolutionary context for monogamy in the Ornate Tinamou, the present study aimed to determine whether this species is genetically monogamous or not and whether males are subjected to cuckoldry. We used polymorphic microsatellite markers to perform a sib-ship and paternity analysis based on samples from eggs and feathers of incubating males from four clutches. Promiscuity was found in all nests and the observed rate of cuckoldry ranged from 0% to 33.3%. These results rebut the consideration of monogamy for the Ornate Tinamou. Thus, the optimal model with polygynous males and polyandrous females prevails as an evolutionary stable strategy among tinamous, even in different social mating systems and despite high levels of cuckoldry.

Species often respond differently to identical environmental changes. Such different responses might be the consequence of varying ecological requirements and tolerances of specialists compared to generalists. We selected two closely related burnet moth species with contrasting ecological requirements: Zygaena carniolica is restricted to semi-natural calcareous grasslands and represents a stenotopic taxon, while Zygaena viciae occurs more widespread at extensively used meadows forming larger metapopulations. To examine different responses to recent habitat fragmentation, we analysed genetic and morphologic variation between populations of these two Zygaenid species in western Germany and adjoining areas in France and Luxembourg. Our data indicate contrasting genetic and morphologic patterns for both species: the genetic differentiation among populations is lower for Z. carniolica (FST: 0.049) compared to Z. viciae (FST: 0.058). The genetic diversity for Z. carniolica is rather low and homogeneously distributed over all populations, while Z. viciae has more alleles, which are unevenly distributed and often restricted to single populations, as shown by GENELAND models. The level of morphologic variance among and within (e.g. asymmetry) individuals is low for Z. carniolica, but much higher for Z. viciae. We conclude that these contrasting patterns mirror opposite responses on recent habitat degradation and associated loss of interconnectivity as a result of habitat destruction and land-use intensification. The data suggest that species like Z. viciae, with (historically) higher abundances suffer even stronger under the recent habitat loss. Morphologic asymmetry and genetic differentiation in the latter species rises, while diversity declines. More specialised taxa occurring in naturally isolated habitats (indicated by Species Distribution Models) feature an originally lower level of genetic diversity, and thus the impact of habitat decline on species’ persistence is less dramatic, as shown for Z. carniolica. Hence, we conclude that past and recent occurrence pattern has to be taken into account when interpreting data in the light of conservation measures.

OCHKIRCHSubmitted: 17 September 2011Accepted: 10 April 2012doi:10.1111/j.1463-6409.2012.00548.xHusemann, M., Namkung, S., Habel, J.C., Danley, P.D. & Hochkirch, A. (2012).Phylogenetic analyses of band-winged grasshoppers (Orthoptera, Acrididae, Oedipodinae)reveal convergence of wing morphology. —Zoologica Scripta, 41, 515–526.Historically, morphological traits have been used to examine the relationships of distantlyrelated taxa with global distributions. The use of such traits, however, may be misleadingand may not provide the resolution needed to distinguish various hypotheses that mayexplain the distribution patterns of widely distributed taxa. One such taxon, the Oedipo-dine grasshoppers, contains two tribes principally defined by wing morphologies: the Bry-odemini have broad wings whereas Sphingonotini are narrow-winged. Through the use ofmorphological features alone, hypotheses concerning the evolution of these tribes cannotbe distinguished. To differentiate hypotheses that may explain the distribution of Oedipo-dines, such as vicariance, natural dispersal and anthropogenic translocation, we used twomitochondrial and three nuclear gene fragments to reconstruct the phylogenetic relation-ships within and between the two tribes, and employed a molecular clock to evaluate thehypotheses of vicariance and dispersal. Our results clearly reject monophyly of the tribesand revealed monophyletic Old and New World clades, which is in agreement with previ-ous molecular studies. The split between both clades was dated at 35 Ma (±12 Ma). Thisclearly rejects the vicariance hypothesis and supports a single invasion via the Beringianland bridge. In addition, our data clearly show that the similarities in wing morphologyused for distinguishing both tribes are the result of at least one convergent event. Ourstudy shows that interpretations of relationships based on the currently accepted taxonomyin the study groups will be error prone. We suggest that future revisions that consider ourfindings are required.Corresponding author: Martin Husemann, Biology Department, Baylor University, One BearPlace #97388, Waco, TX 76798, USA. E-mail: martin_husemann@baylor.eduSuk Namkung and Patrick D. Danley, Biology Department, Baylor University, One Bear Place#97388, Waco, TX 76798, USA. E-mails: Suk_Namkung@baylor.edu; Patrick_Danley@baylor.eduJan C. Habel, Invertebrate Biology, Natural History Museum, Luxembourg 2160, Luxembourg. E-mail: Janchristianhabel@gmx.deAxel Hochkirch, Department of Biogeography, Trier University, D-54286 Trier, Germany. E-mail:hochkirch@uni-trier.de

When low dispersal ability of an organism meets geographical barriers, the evolution of inter- and intraspecific differentiation is often facilitated. In the Atlas massif of North Africa, the genus Buthus splits into several species and diverges into numerous genetic lineages, often following the orographic structures of mountain systems. Such high mountain ranges often act as barriers for species with reduced mobility even on small spatial scales. To study the effect of orographic structures on organisms with low dispersal ability, we collected 61 individuals of the scorpion species Buthus elmoutaouakili at 18 locations around the southwestern foothills of the High Atlas and Antiatlas and in the Sousse valley (western Morocco). We analyzed intraspecific differentiation patterns within this geographically restricted area of about 100 × 50 km using 452 bp of the cytochrome oxidase I mitochondrial gene. We detected 5 distinct genetic lineages. In a second analysis, we added 61 previously published sequences from Buthus species from Europe and North Africa. Using a molecular clock approach, we detected old splits (4-5 Ma) separating the samples from 1) the western High Atlas and north of these mountains, 2) the Sousse valley and adjoining mountain areas, and 3) the southwestern Antiatlas. Further differentiation happened in the first 2 geographical groups about 3 Ma. Thus, the divergence time estimates based on a Bayesian approach support the onset of differentiation into these main clades along the Pliocene (5-2.3 Ma) when climatic oscillations started and a constant global cooling preceded the glacial-interglacial cycles of the Pleistocene. Further genetic splits into parapatric groups are detectable for the Sousse valley main group in the early Pleistocene. The climatic oscillations of the Pliocene and early Pleistocene might have caused repeated range shifts, expansions, and retractions leading to repeated vicariance, hereby producing the hierarchical structure of genetic differentiation in B. elmoutaouakili. A taxonomic revision, including morphological and molecular data, is needed to assess the status of each of these Buthus scorpion lineages.

The intensification of agricultural land use over wide parts of Europe has led to the decline of semi-natural habitats, such as extensively used meadows, with those that remain often being small and isolated. These rapid changes in land use during recent decades have strongly affected populations inhabiting these ecosystems. Increasing habitat deterioration and declining permeability of the surrounding landscape matrix disrupt the gene flow within metapopulations. The burnet moth species Zygaena loti has suffered strongly from recent habitat fragmentation, as reflected by its declining abundance. We have studied its population genetic structure and found a high level of genetic diversity in some of the populations analysed, while others display low genetic diversity and a lack of heterozygosity. Zygaena loti was formerly highly abundant in meadows and along the skirts of forests. However, the species is currently restricted to isolated habitat remnants, which is reflected by the high genetic divergence among populations (F ST: 0.136). Species distribution modelling as well as the spatial examination of panmictic clusters within the study area strongly support a scattered population structure for this species. We suggest that populations with a high level of genetic diversity still represent the former genetic structure of interconnected populations, while populations with low numbers of alleles, high F IS values, and a lack of heterozygosity display the negative effects of reduced interconnectivity. A continuous exchange of individuals is necessary to maintain high genetic variability. Based on these results, we draw the general conclusion that more common taxa with originally large population networks and high genetic diversity suffer stronger from sudden habitat fragmentation than highly specialised species with lower genetic diversity which have persisted in isolated patches for long periods of time.

The climate warming of the postglacial has strongly reduced the distribution of cold-adapted species over most of Central Europe. Such taxa have therefore become extinct over most of the lowlands and shifted to higher altitudes where they have survived to the present day. The lycaenid butterfly Lycaena helle follows this pattern of former widespread distribution and later restriction to mountain areas such as the European middle mountains. We sampled 203 individuals from 10 populations representing six mountain ranges (Pyrenees, Jura, Massif Central, Morvan, Vosges and Ardennes) over the species' western distribution. Allozyme and microsatellite polymorphisms were analysed to study the genetic status of these highly fragmented populations. Both molecular marker systems revealed a strong genetic differentiation among the analysed populations, coinciding with the orographic structure and highly restricted gene flow among them. The large-scale genetic differentiation is more pronounced in allozymes (F CT : 0.326) than in microsatellites (R CT : 0.113), but microsatellites show a higher resolution on the regional scale (R SC: 0.082) compared with allozymes (F SC : n.s.). For both analytical tools, we found private alleles occurring exclusively in a single mountain area. The highly fragmented and isolated occurrence of populations is supported by the distribution pattern of potentially suitable climate suggested by species distribution models. Model projections under two climate warming scenarios predict a decline of climatically suitable areas, which will result in the extinction of most of the populations showing unique genetic characteristics.

Cold-adapted species are thought to have had their largest distribution ranges in central Europe during the glacial periods. Postglacial warming caused severe range shifts of such taxa into higher latitudes and altitudes. We selected the boreomontane butterfly Lycaena helle (Denis & Schiffermuller, 1775) as an example to demonstrate the genetic effects of range changes, and to document the recent status of highly fragmented remnant populations. We analysed five polymorphic microsatellite loci in 1059 individuals sampled at 50 different localities scattered over the European distribution area of the species. Genetic differentiation was strong among the mountain ranges of western Europe, but we did not detect similarly distinct genetic groups following a geographical pattern in the more eastern areas. The Fennoscandian populations form a separate genetic group, and provide evidence for a colonization from southern Finland via northern Scandinavia to south-central Sweden. Species distribution modelling suggests a large extension of the spatial distribution during the last glacial maximum, but highlights strong retractions to a few mountain areas under current conditions. These findings, combined with our genetic data, suggest a more or less continuous distribution of L. helle throughout central Europe at the end of the last ice age. As a consequence of postglacial warming, the species retreated northwards to Fennoscandia and escaped increasing temperatures through altitudinal shifts. Therefore, the species is today restricted to population remnants located at the mountain tops of western Europe, genetically isolated from each other, and evolved into genetically unique entities. Rising temperatures and advancing habitat destruction threaten this wealth of biodiversity. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101, 155-168. ADDITIONAL KEYWORDS: climate change - fragmentation - Lepidoptera - microsatellites - mountains - postglacial relict - range shift - species distribution modelling.

Temporal changes in allele frequencies are often assumed in studies addressing the history of populations affected by different anthropogenic and natural impacts at different time scales. Yet, few studies directly compare the genetic composition of populations over time spans of more than 10 years. Therefore, to test the genetic effects of 15 years of population isolation in the butterfly Lycaena helle, we analysed 472 individuals from 27 samples, of which nine were collected in 1991 and 18 in 2006. Sampling was performed in five mountain regions (Pyrenees, Massif Central, Jura, Vosges and Ardennes). Genetic analyses were performed using five polymorphic microsatellites. Old and new samples of identical or neighbouring populations revealed similar genetic differentiations among these five mountain regions. A comparatively strong genetic differentiation among populations combined with a high amount of private alleles for each mountain area was detected, but mountain-specific alleles were in most cases identical in 1991 and 2006. Nevertheless, the obtained data also indicate moderate changes between 1991 and 2006 in the species genetic structure – genetic differentiation among local populations increased marginally and allele frequencies showed corresponding modifications. A significant decline in genetic diversity was not detectable, and nine private alleles exclusive to a single mountain region were only detected in samples from the year 1991, whereas eleven were only observed in the individuals collected in 2006. These observations might indicate the results of genetic drift within isolated populations.

The sea acts as an effective dispersal barrier for most terrestrial animal species. Narrow sea straits, therefore, often represent areas where species are able to disperse from one land mass to another. In the Mediterranean Sea, the narrowest connecting points between North Africa and Europe are the Strait of Gibraltar and the Strait of Sicily. In the past, climatic oscillations caused changing sea levels and thus influenced the permeability of these sea straits. We analysed the genetic structure of four butterfly species that all occur on both sides of the Strait of Sicily. In all four species, we observed a lack of genetic differentiation between the populations of North Africa and those of Italy. Species distribution models support the strong cohesiveness in that they show a largely continuous glacial distribution over Italy and North Africa. The data obtained reveal that there was a large exchange of individuals between Italy and the eastern Maghreb during the last ice age. This might not only be the case for the species under investigation in the present study, but also might represent a more general pattern for mobile thermophilic western Palearctic species. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99, 818–830.

Agricultural intensification caused a fragmentation of flower-rich extensively used meadows which resulted in the reduction of the abundance of species of these habitats. The abundance pattern and the dispersal behaviour of species influence the connectivity of local populations. In this context exchange rates can be directly measured by mark release recapture (MRR) studies and indirectly by genetic analysis. Both approaches are used in our study in a comparative way. As a test species to investigate the influence of habitat interconnectivity on a local and regional scale, we selected the butterfly Melanargia galathea (Lepidoptera, Nymphalidae), a widely distributed species, which is common on flower-rich meadows in our study area in western Germany. We marked 3,175 individuals of four neighbouring sites in a mark-release-recapture study and analysed 18 allozyme loci for 644 individuals sampled over 17 sites. Only 3.3% of the total genetic variance was found among samples, thus supporting the detected between-patch movements. Both approaches revealed a high exchange rate among local populations. Moderate between- and high within-patch movements were recorded by MRR analysis, dependent on the geographical distance. The two analytical tools showed high estimated effective population sizes for all populations. In the light of conservation biology, the combination of the MRR and allozyme data support the assumption that high dispersal ability and habitat interconnectivity countervail genetic differentiation and enable the maintenance of a high level of genetic differentiation.

Rising temperatures and agricultural changes (intensification and succession on fallow land) during the last few decades have caused a strong decline of moist and cool sites on nutrient-poor grasslands and species depending on these habitats. We tested the effects of habitat deterioration on a local and regional scale in such a species, the highly endangered butterfly Lycaena helle, which was more widely distributed over central Europe during the postglacial period, but has recently become restricted to some remnants. We analysed five polymorphic microsatellite loci in 220 individuals sampled at ten different localities. The study sites in Germany, Luxembourg and Belgium are geographically split into three mountain regions: the Ardennes, the Eifel and the Westerwald; the latter is separated from the other two by the river Rhine. A comparatively high genetic diversity was detected in all local populations and genetic differentiation was found among the Ardennes, the Eifel and the Westerwald (F-CT: 0.084). The genetic differentiation among all populations (F-ST: 0.137) underlines natural and anthropogenic habitat fragmentation. While ongoing gene flow seems to exist among the Eifel populations indicating the only intact metapopulation, a high genetic differentiation in the Ardennes and the Westerwald indicates a disruption of population connectivity. Our genetic data obtained on different spatial scales show the genetic consequence of long-term isolation and should trigger necessary conservation measures at the metapopulation level. Rising temperatures and agricultural changes (intensification and succession on fallow land) during the last few decades have caused a strong decline of moist and cool sites on nutrient-poor grasslands and species depending on these habitats. We tested the effects of habitat deterioration on a local and regional scale in such a species, the highly endangered butterfly Lycaena helle, which was more widely distributed over central Europe during the postglacial period, but has recently become restricted to some remnants. We analysed five polymorphic microsatellite loci in 220 individuals sampled at ten different localities. The study sites in Germany, Luxembourg and Belgium are geographically split into three mountain regions: the Ardennes, the Eifel and the Westerwald; the latter is separated from the other two by the river Rhine. A comparatively high genetic diversity was detected in all local populations and genetic differentiation was found among the Ardennes, the Eifel and the Westerwald (FCT: 0.084). The genetic differentiation among all populations (FST: 0.137) underlines natural and anthropogenic habitat fragmentation. While ongoing gene flow seems to exist among the Eifel populations indicating the only intact metapopulation, a high genetic differentiation in the Ardennes and the Westerwald indicates a disruption of population connectivity. Our genetic data obtained on different spatial scales show the genetic consequence of long-term isolation and should trigger necessary conservation measures at the metapopulation level.

Many studies in population ecology have shown that related species have different dispersal behaviours. Species with sedentary and migratory behaviour exist in butterflies. While the genetic responses to population isolation are well studied, the effects of different dispersal behaviours of species are widely unknown. Therefore, we analysed 19 allozyme loci of two lycaenid butterflies,Cupido minimusas a sedentary butterfly andAricia agestisas a mobile and expansive species. We collected 594 individuals (280 ofC. minimusand 314 ofA. agestis) in a western German study region with adjacent areas in Luxembourg and northeastern France. The genetic differentiation among populations ofA. agestis(FST=3.9%) was lower than inC. minimus(FST=5.6%). Both species built up an isolation-by-distance system, which is more pronounced inA. agestisthan inC. minimus. The genetic diversity inC. minumuspopulations (e.g.Ptot=73.5%) is higher for all analysed parameters than inA. agestis(e.g.Ptot=52.1%). Both species show specific genetic characteristics fitting with their different dispersal behaviours and respective ecological strategies. In the light of conservation genetics, we deduce that highly fragmented populations do not necessarily have a high extinction probability, but this risk depending much more on specific population genetic structures. In the studied species,C. minimuspreserves a complex genetic constitution by high population densities. The patchily distributedA. agestisrepresents less rare alleles, present only in some populations, and holds up genetic diversity by high mobility.

Many studies aim at testing the impact of recent fragmentation on the genetic diversity and connectivity of populations, while some species do exist naturally in fragmented landscapes because of their habitat requirements. Therefore, it is important to look at the genetic signatures of species occurring in naturally fragmented landscapes in order to disentangle the effect of fragmentation from the effect of habitat requirements. We selected two Nymphalid butterflies for this purpose. While Melanargia galathea is a common butterfly in flower-rich meadows, Melitaea aurelia is closely connected to calcareous grasslands, thus being historically fragmented due to its ecological demands. For the analysis of the genetic response on these opposed patterns, we analysed 18 allozyme loci for 789 individuals (399 individuals of M. galathea and 390 individuals of M. aurelia) in a western German study region with adjacent areas in Luxemburg and northeastern France. Both species showed similarly low genetic differentiations among local populations (M. galathea: F ST 3.3%; M. aurelia: F ST 3.6%), both combined with a moderate level of inbreeding. Isolation-by-distance analysis revealed a significant correlation for both species with similar amounts of explained variances (M. galathea: r 2 = 27.8%; M. aurelia: r 2 = 28.5%). Most parameters of genetic diversity were higher in M. galathea than in M. aurelia, but the latter species had a considerably higher amount of rare or locally restricted genes; the differing ecological demands are thus reflected in these differences. Both species thus seem to be genetically well suited to their respective ecological requirements. In the light of conservation genetics, we deduce that highly fragmented populations are not necessarily prone to extinction. The extinction risk might be linked to the life history of an organism and its population genetic structure.

Classical Sanger sequencing is still frequently used to generate sequence data for phylogenetic and phylogeographic inference. In this contribution we present a novel approach to genotype whole mitogenomic haplotypes using Illumina MiSeq reads from indexed amplicons. Our new approach reduces preparation time by multiplexing loci within a single or few PCR reactions and by plate format library construction. The use of paired-end reads allows covering amplicons of about 0.5kb and thus no nebulisation and assembly are necessary. We tested the power and effectiveness of this technique by analysing the mitogenomic diversity of East African white-eye bird species (Zosteropidae), a taxonomically highly diverse and complex species flock found in various ecosystems spread across major parts of Africa. We compare the newly generated mitogenomic data set with published data of three mitochondrial genes for a similar set of populations and taxa. The comparison demonstrates that our new procedure represents a cost effective use of NGS for medium throughput phylogenetic analyses. Using this method, we were able to increase the amount of phylogenetic information significantly, while reducing the costs and effort in the laboratory. The mitogenomic data show a higher resolution than previous studies providing higher support and new insights in the relationships of Zosterops species. Our data suggest to split Z. poliogaster into four distinct species, three of which had previously been proposed: Z. silvanus, Z. mbulensis, Z. kikyuensis and Z. kulalensis. Our approach allows the genotyping of whole mitogenomes for a large number of individuals and thus allows more reliable reconstruction of phylogenetic and phylogeographic relationships - also for non-model organisms.

The habitats of many species are fragmented. Therefore, the survival in a metapopulation depends on the stability of the single populations and the amount of movements between patches. We chose the calcareous grassland specialist butterfly species Polyommatus coridon as a model. As study area, we selected a mosaic-like landscape in Rhineland-Palatinate (western Germany) with several well preserved calcareous grassland fragments. We marked a total of 2,211 individuals during July and August 2003. The overall recapture ratio was 7.1%. The estimated mean butterfly densities over the whole flight season ranged from 52 to 487 individuals per hectare. The within-patch movements were relatively low (13.3%) compared with the between-patch movements (3.2%). Therefore, the metapopulation structure appears to be intact in our study area.

Isolation of Mediterranean species in the southern European peninsulas during the cold glacial phases often resulted in differentiation of several genetic lineages confined to the respective peninsulas. However, whilst there is good genetic evidence for multiple refugia in Iberia, there are only limited data available for the Balkans. Therefore, we wish to examine the hypothesis of a strong genetic structuring within southeastern Europe for the existence of multiple Balkan differentiation centres and/or several leading edges. As a model we use the marbled white butterfly, Melanargia galathea . We studied 18 allozyme loci of 564 individuals from 16 populations distributed over a large part of southeastern Europe. The single populations showed moderately high genetic diversity and no northward decline of genetic diversity was detected. The overall genetic differentiation between populations was considerable ( F ST 7.0%). Cluster analysis discriminated three genetic groups: (i) a western flank in the former Yugoslavia, parts of eastern Austria and Hungary; (ii) an eastern flank with populations from Bulgaria and Romania (south of the southern Carpathians and eastern Carpathians); and (iii) the eastern Carpathian Basin. Hierarchical variance analysis distributed 53% of the variance among populations between these three groups. One sample from the Greek–Bulgarian border clustered within the eastern flank, but showed some tendency towards the eastern Carpathian Basin populations. Two populations from Carinthia clustered together with the eastern Carpathian Basin ones and a population from Styria showed an intermediate genetic composition between the three groups. Most probably, the eastern and the western flank groups are due to postglacial range expansion from the northeastern and the northwestern edges of the glacial differentiation centre (so-called leading edges). The eastern Carpathian Basin group may have resulted from postglacial expansion from northern Greece through valley systems of the central Balkan peninsula, maybe even expanding westwards north of the Balkan mountains reaching some parts of eastern Austria (e.g. Carinthia). Therefore, the Balkanic refugium of M. galathea may or may not have been continuous along the coastal areas of the Mediterranean, but must have been strongly genetically structured.

Aim Three paradigm patterns of post-glacial dispersal are known for terrestrial species in Europe. However, the possibility of a fourth arises with the Italian and the Balkan lineages expanding to central Europe and the Iberian one being trapped by the Pyrenees. We test this hypothesis by analysing the molecular biogeography of the Marbled White butterfly. Location Twelve populations distributed over a major part of the European range of Melanargia galathea and M. lachesis. Methods We studied 18 allozyme loci of 403 individuals from 12 populations. Butterflies were sampled in the field, frozen in liquid nitrogen and stored under these conditions until analysis. We used cellulose acetate plates for allozyme electrophoresis. Results We detected three major genetic lineages within the M. galathea/lachesis complex. The M. lachesis sample from the southern Pyrenees was strongly genetically differentiated from M. galathea (FCT: 0.312). Melanargia galathea splits into two major genetic lineages (FCT: 0.115), which both were found in post-glacially invaded regions. The further differentiation within these lineages was comparably low (FSC: 0.028). The genetic diversity within populations was high compared with other butterfly species. Main conclusions Our findings support the existence of a fourth pattern with only the Iberian lineage not contributing considerably to the post-glacial colonization of central Europe. Preliminary studies in other butterfly species of dry grasslands support the importance of this pattern possibly representing a fourth paradigm. The high genetic diversity within populations might be one reason for the recently observed expansions at the northern distribution limits.

Species with specific habitat requirements often remain within their habitat and are characterized by a very sedentary behaviour. We used radio-tracking during a 3-wk campaign to investigate the home ranges and movements of three East African passerine bird species, all adapted to cloud-forest habitats: montane white-eye Zosterops poliogaster mbuluensis, stripe-cheeked greenbul Andropadus milanjensis and white-starred robin Pogonocichla stellata macarthuri. Individuals were observed in a forest-grassland mosaic on top of Chyulu Hills, south Kenya. Based on 15 individuals with a total of 1041 fixes, we found (1) that all three species show a sedentary behaviour restricted to their home forest patch; (2) least activity during the afternoon; and (3) re-colonization into its home patch after translocation into a nearby forest patch. Our findings underline that forest-specialist bird species of the tropics show high site fidelity despite their potential movement ability.

To understand how landscape characteristics affect gene flow in species with diverging ecological traits, it is important to analyze taxonomically related sympatric species in the same landscape using identical methods. Here, we present such a comparative landscape genetic study involving three closely related Hesperid butterflies of the genus Thymelicus that represent a gradient of diverging ecological traits. We analyzed landscape effects on their gene flow by deriving inter-population connectivity estimates based on different species distribution models (SDMs), which were calculated from multiple landscape parameters. We then used SDM output maps to calculate circuit-theoretic connectivity estimates and statistically compared these estimates to actual genetic differentiation in each species. We based our inferences on two different analytical methods and two metrics of genetic differentiation. Results indicate that land use patterns influence population connectivity in the least mobile specialist T. acteon. In contrast, populations of the highly mobile generalist T. lineola were panmictic, lacking any landscape related effect on genetic differentiation. In the species with ecological traits in between those of the congeners, T. sylvestris, climate has a strong impact on inter-population connectivity. However, the relative importance of different landscape factors for connectivity varies when using different metrics of genetic differentiation in this species. Our results show that closely related species representing a gradient of ecological traits also show genetic structures and landscape genetic relationships that gradually change from a geographical macro- to micro-scale. Thus, the type and magnitude of landscape effects on gene flow can differ strongly even among closely related species inhabiting the same landscape, and depend on their relative degree of specialization. In addition, the use of different genetic differentiation metrics makes it possible to detect recent changes in the relative importance of landscape factors affecting gene flow, which likely change as a result of contemporary habitat alterations. peerReviewed

Many studies on the biogeography of thermophilic and arctic-alpine species were performed during the past. Only little is known about species with intermediate characteristics. We analyzed the molecular biogeography of the butterfly Erebia alberganus (30 populations, representing 1106 individuals), sampled over the Alps, Apennines (Italy), and the Stara Planina (Bulgaria) using allozyme electrophoresis (17 loci). Genetic analyses revealed 3 major splits, with the strongest between the Stara Planina populations and all other populations, and a weaker split between the Alps and the Apennines. Individuals from the Apennines were genetically nested within the Alps group. The Alps cluster was segregated into 3 groups: the Southwestern, Western/Central, and Eastern Alps. The genetic diversities were highest for the Alps populations and significantly lower in the 2 isolates (Apennines, Stara Planina). The remarkable genetic split between Stara Planina and all other populations and the genetic distinctiveness of the former cluster might be interpreted as an ancient colonization event of this Balkan mountain range. The Apennines populations derive from a more recent expansion out of the Southwestern Alps. After surviving the Würm ice age most probably in the central Apennines, accompanied by genetic modification of some of these populations, northward expansion might have started from the western parts of the central Apennines reaching the northern Apennines during the early postglacial. The subtle genetic differentiation found among the Alps populations probably reflects 3 geographically disjunct Würm glacial centers located at the western slopes of the Southwestern Alps, at the southern slopes of the Central Alps, and in the Southeastern Alps.

Mountain species have evolved important genetic differentiation due to past climatic fluctuations. The genetic uniqueness of many of these lineages is now at risk due to global warming. Here, we analyse allozyme polymorphisms of 1306 individuals (36 populations) of the mountain butterfly Erebia manto and perform Species Distribution Models (SDMs). As a consensus of analyses, we obtained six most likely genetic clusters: (i) Pyrenees with Massif Central; (ii) Vosges; (iii-v) Alps including the Slovakian Carpathians; (vi) southern Carpathians. The Vosges population showed the strongest genetic split from all other populations, being almost as strong as the split between E. manto and its sister species Erebia eriphyle. The distinctiveness of the Pyrenees-Massif Central group and of the southern Carpathians group from all other groups is also quite high. All three groups are assumed to have survived more than one full glacial-interglacial cycle close to their current distributions with up-hill and down-slope shifts conforming climatic conditions. In contrast with these well-differentiated groups, the three groups present in the Alps and the Slovakian Carpathians show a much shallower genetic structure and thus also should be of a more recent origin. As predicted by our SDM projections, rising temperatures will strongly impact the distribution of E. manto. While the populations in the Alps are predicted to shrink, the survival of the three lineages present here should not be at risk. The situation of the three other lineages is quite different. All models predict the extinction of the Vosges lineage in the wake of global warming, and also the southern Carpathians and Pyrenees-Massif Central lineages might be at high risk to disappear. Thus, albeit global warming will therefore be unlikely to threaten E. manto as a species, an important proportion of the species' intraspecific differentiation and thus uniqueness might be lost.

The immense biodiversity of the Atlas Mountains in North Africa might be the result of high rates of microallopatry caused by mountain barriers surpassing 4000 meters leading to patchy habitat distributions. We test the influence of geographic structures on the phylogenetic patterns among Buthus scorpions using mtDNA sequences. We sampled 91 individuals of the genus Buthus from 51 locations scattered around the Atlas Mountains (Antiatlas, High Atlas, Middle Atlas and Jebel Sahro). We sequenced 452 bp of the Cytochrome Oxidase I gene which proved to be highly variable within and among Buthus species. Our phylogenetic analysis yielded 12 distinct genetic groups one of which comprised three subgroups mostly in accordance with the orographic structure of the mountain systems. Main clades overlap with each other, while subclades are distributed parapatrically. Geographic structures likely acted as long-term barriers among populations causing restriction of gene flow and allowing for strong genetic differentiation. Thus, genetic structure and geographical distribution of genetic (sub)clusters follow the classical theory of allopatric differentiation where distinct groups evolve without range overlap until reproductive isolation and ecological differentiation has built up. Philopatry and low dispersal ability of Buthus scorpions are the likely causes for the observed strong genetic differentiation at this small geographic scale.

We investigated the extent of poleward shifts in the distribution range of Agonum viridicupreum due to climate change in the western Palaearctic. Species' records were obtained from extensive literature sources as well as from collections, and consistent amateur entomologists' recordings. Within the general geographic range of the species, we analyzed in detail two parts of both, the northern and southern distribution range boundaries: (1 and 2) north-western Germany (leading or high-latitude edge), (3) Israel and (4) southern Italy (rear or low-latitude edge). Temporal changes in the occurrence data of the species indicated a northward shift of the leading edge of a minimum of 100 km within the last 50 to 100 years. In contrast, according to the data gathered, the rear edge has not changed during the last decades. Further studies are needed in order to fully understand the underlying mechanisms of the different behaviour of leading and rear range edges of Agonum viridicupreum in the current context of global change. Despite our incomplete understanding, chronosequences of the occurrence of the given species have the potential to optimize climate niche modelling to predict trends in the distribution range in the future. We investigated the extent of poleward shifts in the distribution range of Agonum viridicupreum due to climate change in the western Palaearctic. Species' records were obtained from extensive literature sources as well as from collections, and consistent amateur entomologists' recordings. Within the general geographic range of the species, we analyzed in detail two parts of both, the northern and southern distribution range boundaries: (1 and 2) north-western Germany (leading or high-latitude edge), (3) Israel and (4) southern Italy (rear or low-latitude edge). Temporal changes in the occurrence data of the species indicated a northward shift of the leading edge of a minimum of 100 km within the last 50 to 100 years. In contrast, according to the data gathered, the rear edge has not changed during the last decades. Further studies are needed in order to fully understand the underlying mechanisms of the different behaviour of leading and rear range edges of Agonum viridicupreum in the current context of global change. Despite our incomplete understanding, chronosequences of the occurrence of the given species have the potential to optimize climate niche modelling to predict trends in the distribution range in the future.

Relict Species: From Past to Future.- Climate and Ecosystems.- The Changing Climate: Past, Present, Future.- Extra-Mediterranean Refugia, Post-Glacial Vegetation History and Area Dynamics in Eastern Central Europe.- Ancient Relicts.- Review: The Dark Side of Relict Species Biology: Cave Animals as Ancient Lineages.- Relicts Within the Genus Complex Astragalus/Oxytropis (Fabaceae), and the Comparison of Diversity by Objective Means.- Relict Populations and Endemic Clades in Palearctic Reptiles: Evolutionary History and Implications for Conservation.- Niche Conservatism among Allopatric Species of the Grasshopper Genus Afrophlaeoba Jago, 1983 in the Eastern Arc Mountains (Tanzania).- Conservation Genetics and Phylogeography of the Threatened Corsican and Barbary Red Deer (Cervus elaphus corsicanus and C. e. barbarus).- Phylogeographic Analyses of a Boreal-Temperate Ectomycorrhizal Basidiomycete, Amanita Muscaria, Suggest Forest Refugia in Alaska During the Last Glacial Maximum.- Glacial Relicts.- Review Refugial Areas and Postglacial Colonizations in the Western Palearctic.- Multiple Glacial Refuges of Unwinged Ground Beetles in Europe: Molecular Data Support Classical Phylogeographic Models.- Postglacial Recolonization of Continental Europe by the Pygmy Shrew (Sorex minutus) Inferred From Mitochondrial and Y Chromosomal DNA Sequences.- Relicts Since Postglacial Times.- Are Disjunct Alpine and Arctic-Alpine Animal and Plant Species in the Western Palearctic Really "Relics of a Cold Past"?.- Linking Genetics and Ecology: Reconstructing the History of Relict Populations of an Endangered Semi-Aquatic Beetle.- Peripheral Relict Populations of Widespread Species Evolutionary Hotspots or Just More of the Same?.- Genetic Differentiation Between and Among Refugia.- Population Genetic Structure of Two Threatened Dragonfly Species (Odonata: Anisoptera) as Revealed by RAPD Analysis.- Review Molecular Methods: Blessing or Curse?.- Conservation of Relict Species.- The EU Habitats Directive and the German Natura 2000 Network of Protected Areas as Tool for Implementing the Conservation of Relict Species.- Ecological Limits Vis-a-vis Changing Climate: Relic Erebia Butterflies in Insular Sudeten Mountains.- Conservation and Management of the Habitats of Two Relict Butterflies in the Belgian Ardenne: Proclossiana eunomia and Lycaena helle.- The Future of Relict Species.- Review Modelling Future Trends of Relict Species.- Conservation of the Grey Bush Cricket Platycleis albopunctata (Orthoptera: Tettigoniidae) Under Differing Habitat Conditions: Implications From an Individual-Based Model.- Is the 'Lost World' Lost? High Endemism of Aphibians and Reptiles on South American Tepuis in a Changing Climate.- Population Genetics and Ecological Niche Modelling Reveal High Fragmentation and Potential Future Extinction of the Endangered Relict Butterfly Lycaena helle.- Relict Species Research: Some Concluding Remarks.- Erratum.- Erratum.

During the post-glacial warming, cold-adapted species shifted their distribution to higher latitudes and altitudes and became widely extinct over the European lowlands. The butterfly Lycaena helle shows this feature, and is currently distributed in highly isolated habitat remnants restricted to higher elevations over Central Europe. We analysed five polymorphic microsatellite loci and applied Climate Envelope Modelling. We detected strong genetic differentiation coinciding with the orographic structures of seven distinct mountain regions. This picture become underlined by deflecting levels of genetic diversity and the presence of private alleles, endemic for each single mountain area. Furthermore, genetic differentiation among populations within these mountain groups were detectable and reveal interrupted geneflow on a regional level. This genetic picture of a fragmented distribution coincides with the obtained pattern of potential suitable habitats given by a Climate Envelope Model. A scenario of further climate warming predicts a loss of the major parts of these areas and rising fragmentation of the remainings. The predicted extinction of some populations will cause the loss of unique alleles, which are recently restricted to the given populations.

Multi-locus monomorphism in microsatellites is practically non-existent, with one notable exception, the island fox (Urocyon littoralis dickeyi) population on San Nicolas island off the coast of southern California, having been called ``the most monomorphic sexually reproducing animal population yet reported''. Here, we present the unprecedented long-term monomorphism in relict populations of the highly endangered Parnassius apollo butterfly, which is protected by CITES and classified as ``threatened'' by the IUCN. The species is disjunctly distributed throughout the western Palaearctic and has occurred in several small remnant populations outside its main distribution area. We screened 78 individuals from 1 such relict area (Mosel valley, Germany) at 16 allozyme and 6 microsatellite loci with the latter known to be polymorphic in this species elsewhere. From the same area, we also genotyped 55 museum specimens sampled from 1895 to 1989 to compare historical and present levels of genetic diversity. However, none of all these temporal populations yielded any polymorphism. Thus, present and historical butterflies were completely monomorphic for the same fixed allele. This is the second study to report multi-locus monomorphism for microsatellites in an animal population and the first one to prove this monomorphism not to be the consequence of recent factors. Possible explanations for our results are a very low long-term effective population size and/or a strong historic bottleneck or founder event. Since the studied population has just recovered from a recent population breakdown (second half of twentieth century) and no signs of inbreeding depression have been detected, natural selection might have purged the population of weakly deleterious alleles, thus rendering it less susceptible to the usual negative corollaries of high levels of homozygosity and low effective population size. Multi-locus monomorphism in microsatellitesis practically non-existent, with one notable exception, theisland fox (Urocyon littoralis dickeyi) population on SanNicolas island off the coast of southern California, havingbeen called ‘‘the most monomorphic sexually reproducinganimal population yet reported’’. Here, we present theunprecedented long-term monomorphism in relict populationsof the highly endangered Parnassius apollo butterfly,which is protected by CITES and classified as ‘‘threatened’’by the IUCN. The species is disjunctly distributedthroughout the western Palaearctic and has occurred inseveral small remnant populations outside its main distributionarea. We screened 78 individuals from 1 such relictarea (Mosel valley, Germany) at 16 allozyme and 6microsatellite loci with the latter known to be polymorphicin this species elsewhere. From the same area, we alsogenotyped 55 museum specimens sampled from 1895 to1989 to compare historical and present levels of geneticdiversity. However, none of all these temporal populationsyielded any polymorphism. Thus, present and historicalbutterflies were completely monomorphic for the samefixed allele. This is the second study to report multi-locusmonomorphism for microsatellites in an animal populationand the first one to prove this monomorphism not to be theconsequence of recent factors. Possible explanations forour results are a very low long-term effective populationsize and/or a strong historic bottleneck or founder event.Since the studied population has just recovered from arecent population breakdown (second half of twentiethcentury) and no signs of inbreeding depression have beendetected, natural selection might have purged the populationof weakly deleterious alleles, thus rendering it lesssusceptible to the usual negative corollaries of high levelsof homozygosity and low effective population size.

The fragmentation of landscapes has an important impact on the conservation of biodiversity, and the genetic diversity is an important factor for a populations viability, influenced by the landscape structure. However, different species with differing ecological demands react rather different on the same landscape pattern. To address this feature, we studied three skipper species with differing habitat requirements (Lulworth Skipper Thymelicus acteon: a habitat specialist with low dispersal ability, Small Skipper Thymelicus sylvestris: a habitat generalist with low dispersal ability, Essex Skipper Thymelicus lineola: a habitat generalist with higher dispersal ability). We analysed 18 allozyme loci for 1,063 individuals in our western German study region with adjoining areas in Luxembourg and north-eastern France. The genetic diversity of all three species were intermediate in comparison with other butterfly species. The FST was relatively high for T. acteon (5.1%), low for T. sylvestris (1.6%) and not significant for T. lineola. Isolation by distance analyses revealed a significant correlation for T. sylvestris explaining 20.3% of its differentiation, but no such structure was found for the two other species. Most likely, the high dispersal ability of T. lineola in comparison with T. sylvestris leads to a more or less panmictic structure and hence impedes isolation by distance. On the other hand, the isolation of the populations of T. acteon seems to be so strict that the populations develop independently. Although no general genetic impoverishing was observed for the endangered T. acteon, small populations had significantly lower genetic diversities than big populations, and therefore the high degree of isolation among populations might threaten its local and regional survival. © 2007 Springer Science+Business Media, Inc.

1. Maculinea alcon, a myrmecophilic, stenotopic lycaenid butterfly is restricted to wet heathlands, bogs, and nutrient-poor hay meadows. Due to intensification of agriculture and decrease of extensively grazed meadows, many suitable habitats have disappeared and the remaining ones are highly fragmented and deteriorated.2. Historical distribution data and a comparison with the present occupation of patches show the decline of this critically endangered butterfly in north-west Germany. Most of the populations in north-west Germany are small and often geographically isolated.3. In summer 2002, two-thirds of 77 investigated potential patches were unoccupied as a result of unsuitable habitat structure and habitat fragmentation.4. Several habitat parameters were highly significantly correlated with the presence of M. alcon, in particular the distribution pattern of the host plant Gentiana pneumonanthe. Furthermore, butterflies were absent from many patches with an incidence probability below 50% with respect to patch size and isolation.5. In the nature reserve Lüneburger Heide, part of the study area, M. alcon populations have been observed since 1995. Typical turnover of local populations could be detected during these years. Extinctions and re-colonisations have stabilised the presence of this species in a metapopulation in this nature reserve.6. These data show the importance of different factors on different spatial levels influencing the presence of this endangered butterfly.

Land-use intensity (LUI) is assumed to impact the genetic structure of organisms. While effects of landscape structure on the genetics of local populations have frequently been analysed, potential effects of variation in LUI on the genetic diversity of local populations have mostly been neglected. In this study, we used six polymorphic microsatellites to analyse the genetic effects of variation in land use in the highly abundant grasshopper Chorthippus parallelus . We sampled a total of 610 individuals at 22 heterogeneous grassland sites in the Hainich-Dün region of Central Germany. For each of these grassland sites we assessed habitat size, LUI (combined index of mowing, grazing and fertilization), and the proportion of grassland adjoining the sampling site and the landscape heterogeneity (the latter two factors within a 500 m buffer zone surrounding each focal site). We found only marginal genetic differentiation among all local populations and no correlation between geographical and genetic distance. Habitat size, LUI and landscape characteristics had only weak effects on most of the parameters of genetic diversity of C. parallelus ; only expected heterozygosity and the grasshopper abundances were affected by interacting effects of LUI, habitat size and landscape characteristics. The lack of any strong relationships between LUI, abundance and the genetic structure might be due to large local populations of the species in the landscape, counteracting local differentiation and potential genetic drift effects.

Six polymorphic microsatellite loci were isolated in the endangered butterfly Lycaena helle. Five of them provided interpretable results. We detected four to 34 alleles per locus in a total of 235 samples (males and females) collected from meadows in the Ardennes-Eifel (Germany, Luxemburg and Belgium) and the Westerwald (Germany). We collected one leg for DNA-extraction as a non-lethal method. The expected heterozygosities ranged from 48.6% to 83.1%, depending on the locus analysed. These markers are currently being used in our studies of the species´ phylogeography over its western Palearctic distribution area and for the analysis of the conservation status of the fragmented populations in Central Europe. Six polymorphic microsatellite loci were isolated in the endangered butterfly Lycaena helle. Five of them provided interpretable results. We detected four to 34 alleles per locus in a total of 235 samples (males and females) collected from meadows in the Ardennes-Eifel (Germany, Luxemburg and Belgium) and the Westerwald (Germany). We collected one leg for DNA-extraction as a non-lethal method. The expected heterozygosities ranged from 48.6% to 83.1%, depending on the locus analysed. These markers are currently being used in our studies of the species´ phylogeography over its western Palearctic distribution area and for the analysis of the conservation status of the fragmented populations in Central Europe.

Climatic oscillations influence the distribution of species in time. Thermophilic species survived the ice ages in refugia around the Mediterranean. Northern Africa is one of the possibly important refugia. In this study we test the genetic differentiation between northern African and European populations, using the marbled white butterfly species complex, Melanargia galathea/M. lachesis, as a model. We studied 18 allozyme loci in 876 individuals from 23 populations representing a major part of Europe (northern Spain to Romania) and the western part of northern Africa (Atlas Mountains). The African populations resemble the European ones in allelic richness; their genetic diversity is higher than in Europe. Cluster analysis discriminated five European genetic groups: M. lachesis, a western European lineage, and three eastern European lineages. However, the African samples did not form a separate cluster within this phenogram, but clustered randomly within the Balkan/southeastern European groups. The genetic differentiation among the African populations (FST 8.8%) was higher than that within any of the European lineages (FST 2.6–5.5%). The high genetic diversity and the relatively strong differentiation of the four African populations sampled in a comparatively limited area of the Atlas Mountains indicate that the most probable origin of M. galathea is northern Africa, with its sibling species, M. lachesis, evolving in parallel in Iberia. Most probably, M. galathea colonised Europe first during the Eem interglacial, some 130 ky ago. Since M. lachesis must have existed on the Iberian peninsula during that period already, M. galathea should have reached Europe via Italy. The genetic differentiation to distinct groups in Europe most probably evolved during the following Wurm glacial period.

Background The glacial-interglacial oscillations caused severe range modifications of biota. Thermophilic species became extinct in the North and survived in southern retreats, e.g. the Mediterranean Basin. These repeated extinction and (re)colonisation events led to long-term isolation and intermixing of populations and thus resulted in strong genetic imprints in many European species therefore being composed of several genetic lineages. To better understand these cycles of repeated expansion and retraction, we selected the Marbled White butterfly Melanargia galathea. Fourty-one populations scattered over Europe and the Maghreb and one population of the sibling taxon M. lachesis were analysed using allozyme electrophoresis. Results We obtained seven distinct lineages applying neighbour joining and STRUCTURE analyses: (i) Morocco, (ii) Tunisia, (iii) Sicily, (iv) Italy and southern France, (v) eastern Balkans extending to Central Europe, (vi) western Balkans with western Carpathian Basin as well as (vii) south-western Alps. The hierarchy of these splits is well matching the chronology of glacial and interglacial cycles since the Günz ice age starting with an initial split between the galathea group in North Africa and the lachesis group in Iberia. These genetic structures were compared with past distribution patterns during the last glacial stage calculated with distribution models. Conclusions Both methods suggest climatically suitable areas in the Maghreb and the southern European peninsulas with distinct refugia during the last glacial period and underpin strong range expansions to the North during the Postglacial. However, the allozyme patterns reveal biogeographical structures not detected by distribution modelling as two distinct refugia in the Maghreb, two or more distinct refugia at the Balkans and a close link between the eastern Maghreb and Sicily. Furthermore, the genetically highly diverse western Maghreb might have acted as source or speciation centre of this taxon, while the eastern, genetically impoverished Maghreb population might result from a relatively recent recolonisation from Europe via Sicily.