Judit Bereczki

Oviposition preferences of Maculinea alcon as influenced by aphid (Aphis gentianae) and fungal (Puccinia gentianae) infestation of larval host plants

Abstract 1. The influence of infestation of the larval host plant Gentiana cruciata on the egg‐laying preferences of the xerophilous ecotype of Alcon Blue butterfly (Maculinea alcon) was studied in a semi‐dry grassland area (Aggtelek Karst Region, Northern Hungary).

Relationships within the Melitaea phoebe species group (Lepidoptera: Nymphalidae): new insights from molecular and morphometric information

The genus Melitaea consists of about 80 species, divided into ten species groups, which are all restricted to the Palaearctic region. The Melitaea phoebe group was defined by Higgins based on morphological characters such as wing pattern and genital structures. According to his interpretation, the M. phoebe group included seven species: M. phoebe, M. sibina, M. scotosia, M. aetherie, M. collina, M. consulis and M. turkmanica. The taxonomy of the phoebe species group has been poorly resolved and recent results on the species composition within the group suggest the need for a re‐evaluation. In this study molecular sequences (5985 bp) including one mitochondrial (COI) and up to six nuclear (CAD, EF‐1α, GAPDH, MDH, RpS5 and wingless) gene regions from 38 specimens of the Melitaea phoebe species group sensu Higgins and some closely related taxa from the Palaearctic region were analysed. The possible evolution of the processus posterior of the male genitalia was also reconstructed based on a shape mapping technique. The analysis of the combined data shows a very clear pattern and almost all relationships are highly supported. Based on the combined Bayesian tree and the shape of the processus posterior of the male genitalia, four main groups are recognised: (i) collina group, (ii) arduinna group, (iii) aetherie group and (iv) phoebe group. The status of M. ornata, M. zagrosi and M. scotosia as species is confirmed, and the results also indicate that M. telona (s.s.) from Israel is a separate species.

Genetic structure of Parnassius mnemosyne (Lepidoptera: Papilionidae) populations in the Carpathian Basin

The pattern of genetic variation in a butterfly species depends on the past history of the given species and also on recent evolutionary processes affecting its populations. The aim of the present study was (i) to analyse the enzyme polymorphism in the Clouded Apollo populations of the Carpathian Basin to reveal the contemporary pattern of their genetic differentiation and (ii) to compare it with an expanded mitochondrial DNA (mtDNA) haplotype network of the SE European populations. Allozyme polymorphism was analysed in 22 populations of four geographic regions: Transdanubian (TM) and North Hungarian Mountains (NM), Körös (KÖR) and Bereg–Apuseni–East Carpathian regions (BEAC). The results of the Bayesian clustering analyses based on allozymes supported the presence of three main genetic lineages in the Carpathian Basin: One was typical for TM, another was characteristic for NM and the third cluster was predominant in KÖR. The populations of BEAC harboured a mixture of two clusters. The mtDNA haplotype network suggested a fairly similar distribution: The peri-Alpine clade together with the West Balkan clade was detected in TM, while the East Balkan clade occurred in NM, partly in TR and in the two eastern regions of the Basin (KÖR and BAEC). The incongruities between the results of the mtDNA and allozyme studies can be explained by the different timescales represented by the two markers. The mtDNA haplotype network provided strong evidence concerning the existence of two Balkan lineages, which probably formed a ‘zone of admixture’ in the Transdanubian and North Hungarian Mountains. The possibility of Last Glacial survival of Parnassius mnemosyne in the Carpathian Basin and the conservation implications of these results are discussed.

Patterns of genetic and taxonomic differentiation in three Melitaea (subg. Mellicta) species (Lepidoptera, Nymphalidae, Nymphalinae)

In conservation genetics the existence of Evolutionarily Significant Units (ESU) is a crucial question in threatened or vulnerable species. It is of particular concern to determine whether different subspecies or ecotypes of a species can be considered as separate ESUs. Some Melitaea subg. Mellicta species (e.g. Mellicta aurelia, Mellicta britomartis) are declining or scarce in Europe. Therefore, the level of genetic differentiation and pattern of genetic variation were surveyed in three Melitaea (subg. Mellicta) species. Their habitat requirements and food plants partly overlap; accordingly they often co-occur in the same habitat. M. britomartis and M. aurelia have one brood per year in Hungary, while Mellicta athalia has a monovoltine and a bivoltine ecotype. The purpose of the study was to estimate the number of genetically differentiated ESUs among these species in the Carpathian basin. Samples were taken from 5 Hungarian regions and a few samples were collected in Transylvania as well. Enzyme polymorphism was studied using polyacrylamide gel electrophoresis. The structure of genetic variation was analysed by F-statistics, AMOVA, PCA and Bayesian clustering method. UPGMA dendrogram was constructed on the basis of Cavalli-Sforza and Edwards chord distances. The three species are clearly differentiated from each other in all statistical analyses. They are evidently different Evolutionary Significant Units. The two ecotypes of M. athalia, however, do not show any genetic differentiation.

Regional pattern of genetic variation in the Eastern Central European populations of Euphydryas maturna (Lepidoptera: Nymphalidae)

The aim of this study was to delineate functional conservation units (FCUs) in Scarce Fritillary Euphydryas maturna (Linnaeus, 1758) populations in eastern Central Europe. We analyzed the level and structure of genetic variation using allozymes as markers in 26 Scarce Fritillary populations originating from nine geographic regions. Considering the reproductive strategy of E. maturna, we assumed that populations are strongly exposed to genetic drift. Accordingly, we hypothesized: (i) A low level of genetic variation within populations, and; (ii) A high level of differentiation among them with little evidence of geographic pattern. The results of this study mostly fulfilled our expectations: (i) Scarce Fritillary populations of eastern Central Europe exhibited a relatively low level of polymorphism at investigated enzyme loci, and; (ii) Comparatively strong differentiation was detected among populations. As opposed to our expectation, however, a more-or-less clear pattern of differentiation was revealed by the results of our Bayesian-clustering analysis. Four genetic regions were distinguished on the basis of their average cluster membership coefficients, specifically Dobrogea (Romania), North and East Hungary (composed of four geographic regions), Central and West Hungary (involving three geographic regions), and Transylvania. Three of these genetic regions were previously distinguished as separate subspecies, comprising E. maturna idunides (Central and West Hungary), E. maturna partiensis (North and East Hungary including Transylvania), and E. maturna opulenta (Dobrogea, Romania). These three putative subspecies can be considered as Scarce Fritillary FCUs in eastern Central Europe.

Isolation and characterisation of 15 microsatellite loci from Lethrus apterus (Coleoptera: Geotrupidae)

Fifteen new microsatellite markers for the beetle Lethrus apterus were developed and tested in 45 specimens from the North Hungarian Mountains. Fourteen of the markers developed were polymorphic, and the number of alleles per locus ranged from two to nine. The observed and expected heterozygosities of the polymorphic markers ranged from 0.178 to 0.578, and 0.201 to 0.698, respectively. One locus showed significant deviation from the Hardy-Weinberg equilibrium, probably due to null alleles. The primers were tested on four other Lethrus species (L. bituberculatus, L. scoparius, L. strymonensis and L. perun) and six other coleopteran species (Copris hispanus, Geotrupes stercorarius, Melolontha melolontha, Onthophagus taurus, Oryctes nasicornis and Protaetia affinis). Thirteen loci showed cross-amplification in the Lethrus species and only three loci could be amplified in some of the six other coleopteran species. The developed markers will be valuable in investigating the population genetic structure, behaviour and reproductive biology of L. apterus.

Insulin/IGF Signaling and Life History Traits in Response to Food Availability and Perceived Density in the Cnidarian Hydra vulgaris.

Insulin/insulin-like growth factor signaling (IIS) is thought to be a central mediator of life history traits, but the generality of its role is not clear. Here, we investigated mRNA expression levels of three insulin-like peptide genes, the insulin-like receptor htk7, as well as several antioxidant genes, and the heat-shock protein hsp70 in the freshwater cnidarian Hydra vulgaris. Hydra polyps were exposed to a combination of different levels of food and perceived population density to manipulate life history traits (asexual reproduction and oxidative stress tolerance). We found that stress tolerance and the rate of asexual reproduction increased with food, and that these two effects were in significant interaction. Exposing animals to high perceived density resulted in increased stress tolerance or reduced reproduction only on lower food levels, but not on high food. The insulin-like receptor htk7 and the antioxidant gene catalase were significantly upregulated in the high density treatments. However, the expression level of insulin-like peptide genes, most antioxidant genes, and hsp70 were not affected by the experimental treatments. The higher expression level of htk7 may suggest that animals maintain a higher level of preparedness for insulin-like ligands at high population densities. However, the lack of difference between food levels suggests that IIS is not involved in regulating asexual reproduction and stress tolerance in hydra, or that its role is more subtle than a simple model of life history regulation would suggest.

Additional polymorphic microsatellite loci for detailed population genetic studies of Lethrus Apterus (Coleoptera: Geotrupidae)

In this study we attempted to isolate 13 additional microsatellites from the beetle Lethrus apterus and then to use them to screen for polymorphisms in 35 specimens collected from the northern Hungarian mountains. The newly developed markers presented in this paper are polymorphic, including between two and seven detected alleles per locus. The observed and expected heterozygosity per locus ranged between 0.057 and 0.686 and between 0.056 and 0.756, respectively, while two loci showed significant deviations from Hardy-Weinberg equilibrium. These deviations can be explained by the presence of null alleles. All loci cross- amplified in at least two out of four congeneric Lethrus species (L. bituberculatus, L. scoparius, L. strymonensis and L. perun). The polymorphic microsatellite markers we present for L. apterus will be valuable for future population genetic studies.

Loss of genetic variation in declining populations of Aricia artaxerxes in Northern Hungary

Rapid decline of population size often gives rise to loss of genetic variation and may result in a reduction in fitness. It is, therefore, essential to detect past demographic changes in populations of endangered species. Aricia artaxerxes occurs in two regions of Northern Hungary: Bükk Mts. and the Aggtelek Karst region. The species prefers short-grass habitats, which are subject to succession. Due to climatic differences between the two regions succession is more rapid in the Bükk plateau resulting in the decline and isolation of A. artaxerxes populations there. In contrast, most populations have remained rather large and stable in the Karst region. This situation provides an excellent possibility to compare the genetic composition of stable and declining populations. A. artaxerxes was collected in four populations of the Bükk Mountains and in ten in the Aggtelek Karst region in 2002 and 2005. We analysed 19 polymorphic allozyme loci in all samples. The results revealed an extremely high level of enzyme polymorphism in A. artaxerxes. Most parameters of polymorphism proved to be significantly higher in the Karst region compared to the Bükk. The shape of the distribution of alleles among the frequency classes indicated the effect of bottleneck in three Bükk and one Karst population. Though heterozygote excess was only significant in one Bükk population a tendency was observed for higher values of standardised heterozygote excess in those populations that had a typical bottleneck distribution of alleles. Our results clearly demonstrated that those populations/subpopulations that compose a network with intense migration are able to maintain their genetic diversity in a long run, while the isolated small populations of the Bükk plateau have lost a sizeable part of their variation.