Molecular barcoding for central-eastern European Crioceris leaf-beetles (Coleoptera: Chrysomelidae)
Abstract Among Crioceris leaf-beetles, the two most widespread species (Crioceris asparagi and C. duodecimpunctata) are serious invasive plant pests, while another two (C. quatuordecimpunctata and C. quinquepunctata) are rare species restricted to steppe-like habitats in Eurasia. The aim of the research was to check the genetic distinctiveness of these four species and develop barcodes for their molecular identification using the mitochondrial Cytochrome Oxidase I (COI) gene and two nuclear markers: Elongation Factor 1-α (EF1-α) and Internal Transcribed Spacer 1 (ITS1). The identification of each species was possible and reliable with the use of COI and ITS1 markers. EF1-α was omitted in an…
Exploring species-level taxonomy in the Cryptocephalus flavipes species complex (Coleoptera: Chrysomelidae)
In insects, morphological species identification is often challenging. The discrimination of closely related species may be hampered when only subtle differences in phenotypic characters or a continuum in their variability are present. This is exemplified in the Cryptocephalus flavipes species complex (Coleoptera; Chrysomelidae) where, until now, the species have been discriminated only by the yellow pattern on frons, pronotum, and epipleurae. In the present study, the phylogeny of the C. flavipes species complex was resolved through a multi-locus sequence approach, and the inclusion in the group of the phenotypically similar Cryptocephalus quadripustulatus Gyllenhal, 1813 was evaluated. Su…
Phylogeographic patterns of steppe species in Eastern Central Europe: a review and the implications for conservation
The phylogeography of species associated with European steppes and extrazonal xeric grasslands is poorly understood. This paper summarizes the results of recent studies on the phylogeography and conservation genetics of animals (20 taxa of beetles, butterflies, reptiles and rodents) and flowering plants (18 taxa) of such, "steppic" habitats in Eastern Central Europe. Most species show a similar phylogeographic pattern: relatively high genetic similarity within regional groups of populations and moderate-to-high genetic distinctiveness of populations from currently isolated regions located in the studied area. This distinctiveness of populations suggests a survival here during glacial maxima…
Conservation genetics of highly isolated populations of the xerothermic beetleCrioceris quatuordecimpunctata(Chrysomelidae)
Xerothermic species are rare and threatened in central and eastern Europe. In light of the continuing loss of steppe-like habitats due to anthropogenic fragmentation and degradation, the evaluation of genetic variation in populations inhabiting them is of immediate importance if appropriate conservation measures are to be undertaken. Here we report on the genetic diversity of the rare leaf beetle Crioceris quatuordecimpunctata, whose populations in central and eastern Europe inhabit highly geographically isolated areas. All of the studied populations (in Poland, Ukraine, and Slovakia) were differentiated at the mitochondrial marker COI. However, with respect to the nuclear marker ITS1, Poli…
Low effective population sizes and limited connectivity inxerothermic beetles: implications for the conservation ofan endangered habita
Fragmentation and isolation may have detrimental effects on the viability of populations. Xerothermic grasslands, which are extra-zonal analogues of steppes, are among Europe's most endangered natural environments. Information about connectivity between the remaining habitat patches is critical for effective conservation planning. However, very little is known about the actual levels of isolation of individual xerothermic patches at the inter-regional and local scale. In this study, 16 microsatellite loci were used to investigate genetic diversity, structuring, effective sizes (Ne), and connectivity among populations of the weevil Centricnemus leucogrammus at various geographic scales. Anal…
Restricted geographic distribution and low genetic distinctiveness of steppic Crioceris quinquepunctata (Coleoptera: Chrysomelidae) populations in Central East Europe
Crioceris quinquepunctata is a European leafbeetle, rare and strictly associated with steppe-like habitats in Central and Eastern Europe. We sampled suitable localities in Central East Europe to determine the current distribution and to verify whether populations isolated by the Carpathian Mountains (within Pannonian and Pontic area) show genetic differentiation. Sequences from the beetle COI and ITSl were amplified and compared. Furthermore, ftsZ and hcpA genes of the endosymbiont Wolbachia were analysed as additional genetic markers. We found only two populations of C. quinquepunctata (in Moravia and Podolian Upland). Unusually low genetic differences between these populations were reveal…
Biogeography and ecology of geographically distant populations of sibling Cryptocephalus leaf beetles
Different populations of two closely related species, Cryptocephalusflavipes and C. bameuli, from western (Alps, Apennines and Pyrenees) and central Europe (Poland, Ukraine and Pannonia) were analysed. On the basis of DNA sequences from two genes, cox1 and ef1-alpha, distinctiveness of both species was confirmed. Nevertheless, possible hybrids were identified in Carpathian mountains. We found a significant genetic differentiation among populations of C. flavipes and C. bameuli from distant regions but a high genetic similarity between populations of C. bameuli from north and south of the Carpathians. Demographic estimates suggest a past population expansion in the case of C. bameuli and a r…
Deep divergence and evidence for translocations between Iranian and European populations of the alfalfa weevil (Coleoptera: Curculionidae) based on mitochondrial DNA
AbstractThe alfalfa weevil (Hypera postica(Gyllenhal): Coleoptera: Curculionidae) is an invasive pest of alfalfa (Medicago sativaLinnaeus; Fabaceae) in the Holarctic region. Across the wide geographic distribution of the species different local adaptions have been observed. Further, several distinct mitochondrial lineages have been discovered, which have been treated as western and Egyptian/eastern strains. However, our knowledge of the biogeography ofH. posticais largely limited to North American and Japanese populations. We sampled the species from four locations in Iran and two countries in Europe (Poland and Czech Republic) and sequenced the mitochondrial COI gene to detect patterns of …
Plant-herbivorous beetle networks : molecular characterization of trophic ecology within a threatened steppic environment
DNA barcoding facilitates many evolutionary and ecological studies, including the examination of the dietary diversity of herbivores. In this study, we present a survey of ecological associations between herbivorous beetles and host plants from seriously threatened European steppic grasslands. We determined host plants for the majority (65%) of steppic leaf beetles (55 species) and weevils (59) known from central Europe using two barcodes (trnL and rbcL) and two sequencing strategies (Sanger for mono/oligophagous species and Illumina for polyphagous taxa). To better understand the ecological associations between steppic beetles and their host plants, we tested the hypothesis that leaf beetl…
Preliminary evidence of the horizontal transmission of Wolbachia between Crioceris leaf beetles (Coleoptera: Chrysomelidae) and their Asparagus host plants
Intracellular bacteria of the genus Wolbachia (alpha-Proteobacteria) are the most widespread endosymbionts of insects. Host infection is usually associated with alterations in reproduction, such as cytoplasmic incompatibility, the induction of parthenogenesis and offspring sex ratio bias: all phenomena that may influence host speciation. In the present study, by using well-established molecular tools, we investigated the presence of Wolbachia in leaf beetles of the genus Crioceris and their host plants, which are various species of Asparagus. Multilocus sequence typing of bacterial genes showed that despite their occurrence in the same habitat and feeding on the same plant, two species of C…
How Hosts Taxonomy, Trophy, and Endosymbionts Shape Microbiome Diversity in Beetles.
Bacterial communities play a crucial role in the biology, ecology, and evolution of multicellular organisms. In this research, the microbiome of 24 selected beetle species representing five families (Carabidae, Staphylinidae, Curculionidae, Chrysomelidae, Scarabaeidae) and three trophic guilds (carnivorous, herbivorous, detrivorous) was examined using 16S rDNA sequencing on the Illumina platform. The aim of the study was to compare diversity within and among species on various levels of organization, including evaluation of the impact of endosymbiotic bacteria. Collected data showed that beetles possess various bacterial communities and that microbiota of individuals of particular species h…
The Impact of Environmental Conditions on Efficiency of Host Plant DNA Barcoding for Polyphagous Beetles.
Recently, several papers were published dealing with host plant identification for selected species of insects, including beetles. These studies took advantage of the DNA barcoding approach and generally showed that it is possible to identify diet composition from plant DNA present in insect guts. However, none of these studies considered how the impact of environmental conditions affected the likelihood of insect feeding and, therefore, the presence of host plant DNA that could be amplified and sequenced. In the present study, individuals of the polyphagous weevil Centricnemus leucogrammus (Germar, 1824) (Curculionidae: Entiminae) were used to test the hypothesis that harsh environmental c…
Using host species traits to understand the Wolbachia infection distribution across terrestrial beetles.
AbstractKnowledge of Wolbachia prevalence with respect to its hosts is restricted mainly to taxonomic/phylogenetic context. In contrast, relations between infection and most host’s ecological and biological traits are poorly understood. This study aimed to elaborate on relations between bacteria and its beetle hosts in taxonomic and the ecological contexts. In particular, the goal is to verify which ecological and biological traits of beetles could cause them to be prone to be infected. Verification of Wolbachia infection status across 297 beetle taxa showed that approximately 27% of taxa are infected by supergroups A and B. Only minor support for coevolution between bacteria and its beetle…
The Origin of Isolated Populations of the Mountain Weevil, Liparus glabrirostris—The Flagship Species for Riparian Habitats
AbstractLiparus glabrirostris is one of the largest European weevils, and it has been recently proposed as the flagship species for threatened riparian habitats in the mountains. For effective conservation of its populations (and associated habitats), it is crucial to understand its history, contemporary distribution, genetic diversity and predict changes in the range, including its highly isolated populations on the Baltic coast. Here, we examined numerous populations of L. glabrirostris across almost the entire species range using phylogeography and species niche modeling (SNM) approaches. Analyses of mtDNA and nucDNA markers revealed the existence of 2 major mitochondrial lineages genera…
Conservation genetics of endangered leaf-beetle Cheilotoma musciformis populations in Poland
Steppe-like habitats in Europe are seriously threatened as a result of fragmentation and anthropogenic degradation, at least in western and central parts. Considering the dramatic loss of steppe-like habitats, the evaluation of genetic variation in populations of steppe species is of immediate importance if appropriate conservation measures are to be undertaken. In this paper, we examine the genetic diversity of the highly endangered populations of the leaf-beetle Cheilotoma musciformis, which inhabits only a limited area in south-central Poland, which is geographically isolated from the continuous range of this species. Both mitochondrial and nuclear markers show that the Polish population…
Selective sweep ofWolbachiaand parthenogenetic host genomes - the example of the weevilEusomus ovulum
Most parthenogenetic weevil species are postulated to have originated via hybridization, but Wolbachia has also been speculated to play a role via the induction of parthenogenesis. Here, we examine the molecular diversity of Wolbachia and parthenogenetic host genomes. The host species studied here, Eusomus ovulum, is known to be exclusively parthenogenetic and triploid. The E. ovulum populations that we examined had a low genetic diversity of mitochondrial (cytochrome oxidase I gene) and nuclear markers (internal transcribed spacer 2 and elongation factor 1-α gene), and they all were infected by only single bacteria strains (genotyped for five genes according to the multilocus sequence typi…
Data from: Plant – herbivorous beetle networks: molecular characterization of trophic ecology within a threatened steppic environment
DNA barcoding facilitates many evolutionary and ecological studies, including the examination of the dietary diversity of herbivores. In this study, we present a survey of ecological associations between herbivorous beetles and host plants from seriously threatened European steppic grasslands. We determined host plants for the majority (65%) of steppic leaf beetles (55 species) and weevils (59) known from central Europe using two barcodes (trnL and rbcL) and two sequencing strategies (Sanger for mono/oligophagous species and Illumina for polyphagous taxa). To better understand the ecological associations between steppic beetles and their host plants, we tested the hypothesis that leaf beetl…