Alfred Seitz

The influence of land use on the genetic structure of populations of the common frog Rana temporaria

In order to find out the influence of land use and topographic distance on the genetic structure of populations of the common frog Rana temporaria L. in the Saar-Palatinate lowlands (Federal Republic of Germany), tissue of larvae was examined by means of horizontal starch gel electrophoresis. A total of 24 loci coding for 14 different enzymes were studied. Genotype frequencies, allele frequencies and mean heterozygosity were calculated, and genetic distances using Neis formula. Strong deviations from the Hardy-Weinberg equilibrium were found; the degree of homozygosity was higher than expected. Separation by highways reduced average heterozygosity as well as genetic polymorphism of local populations. One area surrounded by roads had high genetic distances to other sampling stations. A multiple regression analysis showed that motorways and railways have a significant (p = 0·03) barrier effect on frog populations within 3–4 km. Meadowland apparently enabled individual exchange in a range between 2 and 7 km. Consequences for the design of biotope systems are discussed.

Breeding phenology and larval distribution of amphibians in a Mediterranean pond network with unpredictable hydrology

Understanding spatial and temporal breeding patterns in Mediterranean amphibian communities is urgent considering the rate of habitat loss. Breeding phenology and breeding habitat selection by amphibians were analysed through the monthly occurrence of larvae in a mosaic of 198 Mediterranean temporary ponds during three years. A generalized linear model (GLM) coupled with principal component analysis showed that, for almost all species, occurrence was significantly positively correlated to pond depth. In addition, pond openness negatively affected the presence of some species. Temporal breeding patterns varied among species. Some species exhibited flexibility in their breeding date (Pelobates cultripes, Pelodytes punctatus, Hyla meridionalis, Rana perezi), while others did not (Triturus marmoratus, Triturus helveticus, Bufo calamita). When faced with inter-annual hydrological variability, the first group had a more constant breeding success than the second. Variable hydrological conditions caused differential larval occurrence of species between years. These fluctuations might favour long-term persistence of the whole amphibian community. We finally discuss the implications of our results for the management of amphibian habitats in the Mediterranean region.

Palaeoclimatic changes explain Anatolian mountain frog evolution: a test for alternating vicariance and dispersal events

Holarctic biodiversity has been influenced by climatic fluctuations since the Pliocene. Asia Minor was one of the major corridors for postglacial invasions in the Palearctic. Today this area is characterized by an extraordinarily rich fauna with close affiliation to European, Asian and Indo‐African biota. However, exact scenarios of range expansion and contraction are lacking. Using a phylogeographical approach we (i) identify monophyletic lineages among Anatolian mountain frogs and (ii) derive a spatio‐temporal hypothesis for the invasion process in Anatolia. We sequenced 540 bp of the mitochondrial 16S rRNA gene from 40 populations of mountain frogs from Anatolia, the Elburz Mountains and the Caucasus. Our samples comprise all known species and subspecies: Rana macrocnemis macrocnemis, R. m. tavasensis, R. m. pseudodalmatina, R. camerani and R. holtzi. They include the type localities of four of these taxa. We used a nested clade analysis (NCA) to infer historical and recurrent events that account for the observed geographical distribution of haplotypes. None of the extant species is monophyletic. Based on a molecular clock calibration using homologous sequences of Western Palearctic water frogs of the same genus, we estimated that a basic radiation into three lineages c. 2 Mya was followed by several dispersal and fragmentation events. The geographical distribution of resident and widespread haplotypes allows us to infer and date scenarios of range expansion and fragmentation that are aligned with dramatic climatic oscillations that have occurred during the last 600 000 years. Consequently, Pliocene and Pleistocene climatic oscillations triggered the evolution of Anatolian mountain frogs through an interplay of vicariance and dispersal events.

The role of swarming sites for maintaining gene flow in the brown long-eared bat (Plecotus auritus).

Bat-swarming sites where thousands of individuals meet in late summer were recently proposed as ‘hot spots’ for gene flow among populations. If, due to female philopatry, nursery colonies are genetically differentiated, and if males and females of different colonies meet at swarming sites, then we would expect lower differentiation of maternally inherited genetic markers among swarming sites and higher genetic diversity within. To test these predictions, we compared genetic variance from three swarming sites to 14 nursery colonies. We analysed biparentally (five nuclear and one sex-linked microsatellite loci) and maternally (mitochondrial D-loop, 550 bp) inherited molecular markers. Three mtDNA D-loop haplolineages that were strictly separated at nursery colonies were mixed at swarming sites. As predicted by the ‘extra colony-mating hypothesis’, genetic variance among swarming sites (VST) for the D-loop drastically decreased compared to the nursery population genetic variance (VPT) (31 and 60%, respectively), and genetic diversity increased at swarming sites. Relatedness was significant at nursery colonies but not at swarming sites, and colony relatedness of juveniles to females was positive but not so to males. This suggests a breakdown of colony borders at swarming sites. Although there is behavioural and physiological evidence for sexual interaction at swarming sites, this does not explain why mating continues throughout the winter. We therefore propose that autumn roaming bats meet at swarming sites across colonies to start mating and, in addition, to renew information about suitable hibernacula.

Influence of habitat fragmentation on the genetic structure of Polyommatus coridon (Lepidoptera: Lycaenidae): implications for conservation

Conservation biologists pay increasing attention to the interdependence between habitat structure and genetic patterns of species and to the importance of conserving genetic diversity. A suitable model species for such a study is Polyommatus coridon, a butterfly of dry calcareous grassland. We studied the population genetic structure of 22 western German populations of this species (874 individuals). We observed no congruence between the observed genetic structure and the geographic arrangement of the studied populations. Genetic diversity was slightly higher in large populations than in small populations. Differentiation among populations was weak or even missing. Populations of an area with high habitat density showed no significant differentiation, whereas areas with low habitat densities showed significant genetic differentiation among populations. At present, hierarchical variance analysis revealed no differentiation between four distinguished limestone areas. The combination of all analytical results allow the definition of management units for P. coridon. Conservation measures are proposed from which a large number of rarer species occurring together with P. coridon will benefit.

Invasion biology and host specificity of the grapevine yellows disease vector Hyalesthes obsoletus in Europe

Within the past 10 years, the yellows disease ‘bois noir’ (BN) has become one of the commercially most important diseases of grapevine [Vitis vinifera L. (Vitaceae)] in Europe. Infection pressure is caused by phytoplasmas of the stolbur 16SrXII‐A group that are transmitted by a planthopper vector, Hyalesthes obsoletus Signoret (Homoptera: Auchenorrhyncha). Infestation happens as an accidental side‐effect of the feeding behaviour of the vector, as vector and pathogen proliferation is dependent on other plants. In Germany, the increase of BN is correlated with the use of a new host plant by the vector, increase in abundance of the vector on the new host plant, and dissemination of host plant‐specific pathogen strains. In this article, we investigate geographic and host‐associated range expansion of the vector. We test whether host‐plant utilization in Germany, hence the increase in BN, is related to genetic host races of the vector and, if so, whether these have evolved locally or have immigrated from southern populations that traditionally use the new host plant. The genetic population analysis demonstrates a recent expansion and circum‐alpine invasion of H. obsoletus into German and northern French wine‐growing regions, which coincides with the emergence of BN. No H. obsoletus mitochondrial DNA haplotype host‐plant affiliation was found, implying that the ability to use alternative host plants is genetically intrinsic to H. obsoletus. However, subtle yet significant random amplified polymorphic DNA (RAPD) genetic differentiation was found among host plant populations. When combined, these results suggest that a geographic range expansion of H. obsoletus only partly explains the increase of BN, and that interactions with host plants also occur. Further possible beneficial factors to H. obsoletus, such as temperature increase and phytoplasma interactions, are discussed.

Effects of habitat fragmentation on the genetic structure of the monophagous butterfly Polyommatus coridon along its northern range margin

Population genetic patterns of species at their range margin have important implications for species conservation. We performed allozyme electrophoresis of 19 loci to investigate patterns of the genetic structure of 17 populations (538 individuals) of the butterfly Polyommatus coridon, a monophagous habitat specialist with a patchy distribution. The butterfly and its larval food plant Hippocrepis comosa reach their northern distribution margin in the study region (southern Lower Saxony, Germany). Butterfly population size increased with host plant population size. The genetic differentiation between populations was low but significant (FST = 0.013). No isolation‐by‐distance was found. Hierarchical F‐statistics revealed significant differentiation between a western and an eastern subregion, separated by a river valley. The combination of genetic and ecological data sets revealed that the expected heterozygosity (mean: 18.5%) decreased with increasing distance to the nearest P. coridon population. The population size of P. coridon and the size of larval food plant population had no effect on the genetic diversity. The genetic diversity of edge populations of P. coridon was reduced compared to populations from the centre of its distribution. This might be explained by (i) an increasing habitat fragmentation towards the edge of the distribution range and/or (ii) a general reduction of genetic variability towards the northern edge of its distribution.

Forests as dispersal barriers for Erebia medusa (Nymphalidae, Lepidoptera)

Summary The nymphalid butterfly Erebia medusa is a typical species of meadows that are poor in nitrogen and fallow land. In forests, it is only found on clearings with meadow-like structures. However, it is missing on wind exposed places, too. 239 imagoes of E. medusa were sampled in the Aggtelek Karst region (north-eastern Hungary) at six different localities at the end of May 1997. The sample sizes varied between 33 and 46. The studied karst area has a typical forest steppe vegetation and is characterised by a mosaic-like distribution of meadows and forests. The sampled butterflies were analysed by allozyme electrophoresis. The level of polymorphism was significantly higher in the samples than the mean of the populations on European scale. Significant genetic differentiation was found between populations but the level of differentiation was low. Genetic distances varied between 0.013 and 0.020 with a mean of 0.017 (± 0.002 S.D.). The estimated F ST value was about 0.005 (± 0.005 S.D.). An isolation by distance analysis showed no significant correlation between geographical and genetic distances (p > 0.05; r = 0.48). Contrarily, a significant correlation between genetic distances and the minimum forest distances between sampling sites was found (p E. medusa seem to be of little influence on the formation of local genetic structuring, whereas the distribution of forests which might act as migration barriers for this species seems to be an important factor.

Population genetic structure of the butterfly Melitaea didyma (Nymphalidae) along a northern distribution range border

The population genetic structure of the butterfly Melitaea didyma was studied along the northern distribution range border in Central Germany by means of allozyme electrophoresis. Individuals were sampled from a total of 21 habitat patches from four regions, and two provinces. Sampling was designed to estimate local vs. regional differentiation. High levels of variability were found, He= 0.14‐0.21. The mean expected sample heterozygosity from one region, Mosel, was significantly lower than from the Hammelburg region, He= 0.17 and 0.19, respectively. Two hierarchical levels of genetic differentiation were found. Within regions individuals sampled from different patches behaved as belonging to one population with high levels of gene flow (Hammelburg FST= 0.015, Mosel FST= 0.044), though local isolation barriers did create a substructuring of these populations. The inbreeding coefficients, FIS, were constant over all sample levels, suggesting a similar distribution of habitat patches within regions. Between regions gene flow was limited. An isolation by distance analysis indicated that the hierarchical structure, at the provincial level, may be breaking down due to isolation of regional populations. A more general observation was that the sampling design may greatly have influenced the estimation of genetic differentiation. Depending on which samples were included, overall FST estimates ranged from 0.059‐0.090.

Growth cycle of the marbled newt ( Triturus marmoratus ) in the Mediterranean region assessed by skeletochronology

We studied growth patterns within a marbled newt population (Triturus marmoratus, Urodela) at the southeastern limit of its geographical range by skeletochronology of the phalanges and humeri. Seasonal and annual changes in growth rates were assessed through the analyses of 56 females, 66 males and 34 juveniles sampled in different seasons and years. Age and growth cycles were estimated by cross sections of the diaphyseal region of the bone. These analyses revealed a clear line of metamorphosis caused by the metamorphic process itself and the transition to the terrestrial environment. A line of arrested growth (LAG) was observed in winter in both juveniles and adults. Another LAG was recorded in summer among adults. These growth patterns corroborated field observations on life-history traits of the population under study, but differ from those reported for the marbled newt in other environments.