Genetic constitution of parthenogenetic form of Polydrusus inustus (Coleoptera: Curculionidae) : hints of hybrid origin and recombinations

Asexual reproduction among insects may arise spontaneously, as a result of infectious agents or hybridization between phylogenetically distinct organisms. Polyploidy, which may be the result of auto- or allopoliploidization (also associated with hybridization), is often connected with parthenogenesis. However, all studied parthenogens of weevils were apomictic, rare meiotic stages have been observed in a few of them what could suggests in their genomes besides the mutational changeability also recombination may occurs. We studied the level of heterozygosity in Internal Transcribed Spacer 2 sequences, and congruence of mitochondrial (cytochrome B) and nuclear genealogies among individuals of parthenogenetic form of Polydrusus inustus. High frequency (87.0%) of heterozygotes in ITS2 sequences may indicate a hybrid origin of the parthenogen. It is possible that this parthenogenetic form arose via a hybridization event between phylogenetically distinct lineages of bisexual P. inustus, known only from Georgia and Turkey, or between this species and one of the closely relatives from the subgenus Scythodrusus. Although result of partition- homogeneity test did not show significant incongruence of mitochondrial and nuclear genealogies, in the network was detected one loop. Most probable explanations for existence of that loop in parthenogenetic form and for origin of rare homozygotes, which were detected in ITS2 sequences, is gene conversion. It is supported by recently cytogenetic studies indicating rare meiotic stages during the oogenesis of triploids of P. inustus

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 revealed, which is in contrast to previous studies on other steppe beetles. The reasons for such low diversity are speculative and probably related to recent natural expansion or man-made translocation of C. quinquepunctata

Habitat displacement effect between two competing owl species in fragmented forests

Many owl species use the same nesting and food resources, which causes strong interspecific competition and spatio-temporal niche separation. We made use of a recent colonisation of Ural Owls (Strix uralensis) in southern Poland to compare habitat preferences of Tawny Owls (Strix aluco) allopatry and sympatry with Ural Owls. We investigated spatial niche segregation of Ural Owl and the Tawny Owl in sympatry and compared habitat preferences of Tawny Owls breeding in allopatry and sympatry. Tawny Owls breeding in sympatry with Ural Owls occupied forests with higher canopy compactness, sites located closer to forest border and to built-up areas, as well as stands with a higher share of fir and spruce and a lower share of beech as compared to sites occupied by Ural Owls. Allopatric Tawny Owls occupied sites with lower canopy compactness and bred at sites located further from forest borders and in stands with lower share of fir and spruce and a higher share of deciduous as compared to sympatric Tawny Owls. As Ural owls are dominant in relation to Tawny Owls, this indicates that the presence of Ural Owls prevents Tawny Owls from occupying deciduous-dominated and old stands located in forest interior areas, far from buildings and forest edges. The results support habitat displacement between the two species when breeding in sympatry. We also show that protection of large forest patches is crucial for the Ural Owl, a species still rare in central Europe, while small patches are occupied by the abundant Tawny Owl

Development and characterization of microsatellite loci in the Centricnemus leucogrammus weevil

Centricnemus leucogrammus is a weevil characteristic of European xerothermic habitats and steppes. The species was probably more widespread during the Pleistocene glaciations, while its current distribution is limited to “warm-stage refugia.” It may be regarded as a typical representative of flightless xerothermophilous beetles. Previous studies concentrated on its genetic variation using mitochondrial genes. Here, we identified, tested and characterized 24 polymorphic microsatellite loci with the use of 454 sequencing of microsatellite enriched genomic libraries. The new set of loci will be used in studies on the population structure of this weevil and may provide valuable information for its conservation

A three-marker DNA barcoding approach for ecological studies of xerothermic plants and herbivorous insects from central Europe

The DNA barcoding technique developed for species identification has recently been adapted for ecological studies (e.g. host plant identification). Comprehensive barcode databases, covering most species inhabiting areas, habitats or communities of interest are essential for reliable and efficient identification of plants. Here we present a three-barcode (plastid rbcL and matK genes and the trnL intron) database for xerothermic plant species from central Europe. About 85% of the xerothermic plant species (126 out of c. 150) known to be associated with xerothermic habitats were collected and barcoded. The database contains barcodes for 117 (rbcL and trnL) and 96 (matK) species. Interspecific nucleotide distances were in the ranges 0–17.9% (0–3.2% within genera) for rbcL, 0–44.4% (0–3.1%) for trnL and 0–52.5% (0–10.9%) for matK. Blast-searching of each sequence in the database against the entire database showed that species-level identification is possible for 89.6% (rbcL), 98.4% (trnL) and 96.4% (matK) of examined plant species. The utility of the presented database for identification of host plants was demonstrated using two insect species associated with xerothermic habitats: the oligophagous leaf-beetle Cheilotoma musciformis (for which two host plants in Fabaceae were identified) and the polyphagous weevil Polydrusus inustus (which was found to feed on 14 host plants, mostly Rosaceae, Asteraceae and Fabaceae). The developed database will be useful in various applications, including biodiversity, phylogeography, conservation and ecolog

The relationships within the Chaitophorinae and Drepanosiphinae (Hemiptera, Aphididae) inferred from molecular-based phylogeny and comprehensive morphological data

The Chaitophorinae is a bionomically diverse Holarctic subfamily of Aphididae. The current classification includes two tribes: the Chaitophorini associated with deciduous trees and shrubs, and Siphini that feed on monocotyledonous plants. We present the first phylogenetic hypothesis for the subfamily, based on molecular and morphological datasets. Molecular analyses were based on the mitochondrial gene cytochrome oxidase subunit I (COI) and the nuclear gene elongation factor-1α (EF-1α). Phylogenetic inferences were obtained individually on each of genes and joined alignments using Bayesian inference (BI) and Maximum likelihood (ML). In phylogenetic trees reconstructed on the basis of nuclear and mitochondrial genes as well as a morphological dataset, the monophyly of Siphini and the genus Chaitophorus was supported. Periphyllus forms independent lineages from Chaitophorus and Siphini. Within this genus two clades comprising European and Asiatic species, respectively, were indicated. Concerning relationships within the subfamily, EF-1α and joined COI and EF-1α genes analysis strongly supports the hypothesis that Chaitophorini do not form a monophyletic clade. Periphyllus is a sister group to a clade containing Chaitophorus and Siphini. The Asiatic unit of Periphyllus also includes Trichaitophorus koyaensis. The analysis of morphological dataset under equally weighted parsimony also supports the view that Chaitophorini is an artificial taxon, as Lambersaphis pruinosae and Pseudopterocomma hughi, both traditionally included in the Chaitophorini, formed independent lineages. COI analyses support consistent groups within the subfamily, but relationships between groups are poorly resolved. These analyses were extended to include the species of closely related and phylogenetically unstudied subfamily Drepanosiphinae, which produced congruent results. Genera Drepanosiphum and Depanaphis are monophyletic and sister. The position of Yamatocallis tokyoensis differs in the molecular and morphological analyses, i.e. it is either an independent lineage (EF-1α, COI, joined COI and EF-1α genes) or is nested inside this unit (morphology). Our data also support separation of Chaitophorinae from Drepanosiphinae

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 populations are distinct from Slovakian and Ukrainian ones. These regional populations should be considered independent conservation units. On the other hand, very little (mtDNA) or no (nuclear DNA) diversity has been found among the Polish subpopulations. This leads to the conclusion that this species has gone through a strong bottleneck leading to a drastic reduction in its genetic diversity prior to the establishment of present-day populations. Host plants have been identified for this species using barcodes, and the only hosts for the Polish and Ukrainian samples are sainfoins Onobrychis spp. while for the Slovakian sample it is either Dorycnium pentaphyllum or Lotus spp. (all Fabaceae). All of these data can be very valuable for the conservation of C. musciformis populations (e.g. for reintroductions)

The secondary contact zone of phylogenetic lineages of the Philaenus spumarius (Hemiptera : Aphrophoridae) : an example of incomplete allopatric speciation

Previous studies on the phylogeography of the meadow spittlebug Philaenus spumarius (L.) (Hemiptera: Aphrophoridae) suggest the existence of a contact zone of its main phylogenetic lineages along mountain chains in Europe and western Asia. This study presents a detailed examination of the population genetics of P. spumarius within the Carpathian Mountains. The main objective was to determine whether the populations inhabiting that area consist of individuals belonging to different genetic units and whether the observed pattern could be an example of secondary contact zone which formed after incomplete allopatric speciation. Specimens from six transects across the Carpathian arc were examined. The mitochondrial phylogeography of the meadow spittlebug in the examined area clearly shows that individuals from both main clades meet and mix there. Representatives of all three main EF1-alpha clades were also found. The present distribution of the main clades with a zone of overlap along the mountain ranges may suggest that these phylogenetic lineages form a young hybrid zone. Moreover, a limited number of individuals were shown to possess heteroplasmic mitochondrial DNA, which gives additional support to intraspecific hybridization. P. spumarius could be used in future work as an excellent model species in investigating population genetics, intraspecific hybridization, and speciation in progress

Phylogeographic patterns of steppe species in Eastern Central Europe: a review and the implications for conservation

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