Miłosz A. Mazur

Molecular barcoding for central-eastern European Crioceris leaf-beetles (Coleoptera: Chrysomelidae)

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 analyses due to its high level of heterozygosity (presence of multiple PCR products). C. duodecimpunctata and C. quatuordecimpunctata were shown to be sister taxa, but the similar genetic distances between all of the species indicate that these species originated almost simultaneously from a common ancestor. Identification of two separate clades in populations of C. quatuordecimpunctata suggested that the clades are isolated and can be considered as separate conservation units.

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 Cheilotomamusciformis, 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 Dorycniumpentaphyllum or Lotus spp. (all Fabaceae). All of these data can be very valuable for the conservation of C.musciformis populations (e.g. for reintroductions).

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 beetles and weevils differ in food selection as a result of their phylogenetic relations (within genera and between families) and interactions with host plants. We found 224 links between the beetles and the plants. Beetles belonging to seven genera feed on the same or related plants. Their preferences were probably inherited from common ancestors and/or resulted from the host plants chemistry. Beetles from four genera feed on different plants, possibly reducing intrageneric competition and possibly due to an adaptation to different plant chemical defences. We found significant correlations between the numbers of leaf beetle and weevil species feeding on particular plants for polyphagous taxa, but not for nonpolyphagous beetles. Finally, we found that the previous identifications of host plants based on direct observations are generally concordant with host plant barcoding from insect gut. Our results expand basic knowledge about the trophic relations of steppic beetles and plants and are immediately useful for conservation purposes.

Selective sweep of Wolbachia and parthenogenetic host genomes – the example of the weevil Eusomus 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 typing system). We found significant signs of linkage disequilibrium and a lack of recombination amongst all of the examined genomes (bacteria and host), which strongly indicates a selective sweep. The lack of heterozygosity in host nuclear genes, missing bisexual populations and selective sweep between the parthenogenetic host and bacteria genomes suggest that parthenogenesis in this species could have originated as a result of infection rather than hybridization. However, the finding that highly similar Wolbachia strains are also present in other parthenogenetic weevils from the same habitat suggests the opposite scenario: bacteria may have infected the already parthenogenetic lineage and taken advantage of the hosts unisexual reproduction.

The Impact of Environmental Conditions on Efficiency of Host Plant DNA Barcoding for Polyphagous Beetles

ABSTRACT 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 conditions limited its feeding activity. The diet of 50 specimens collected during favourable conditions in the middle of the species reproductive period was compared against the diet of 50 specimens collected during harsh environmental conditions. Results clearly showed that almost no weevils fed during rainy and cold conditions and only a minority of individuals (20%) fed during the drought condition (on drought-resistant plants). It is important to consider such factors in any studies dealing with host plant identification and feeding behaviour. Results of ecological studies could lead to erroneous conclusions, e.g., underestimation of number and composition of host plants in the diet of studies species.

The occurrence of Tenebrionidae (Coleoptera) in Poland based on the largest national museum collections

The paper presents the results of a study on the tenebrionid beetles (more than 10,500 specimens belonging to 78 species) collected in Poland and preserved in the following national zoological collections: Institute of Systematics and Evolution of Animals PAS, Cracow (ISEA), Museum and Institute of Zoology PAS, Warsaw (MIZ) and Upper Silesian Museum, Bytom (USMB). The occurrence of 14 rarely recorded species is confirmed. The presence of 14 studied species on the „Polish Red List of Endangered Animals” and „Polish Red Data Book of Animals” is recorded. The common species, Isomira murina (L.), as a senior synonym of I. semiflava (KUST.), should be excluded from the „Polish Red List of Endangered Animals”. Two species should be excluded from Polish fauna because of an erroneous determination: Pedi- nus helopioides AHR. as Crypticus quisquilius (L.), and Isomira icteropa (KUST.) as Isomira murina (L.). New distributional data on the occurrence on Catalogus faunae Poloniae regions is given for 44 species. Centorus elongatus (HERBST), Cryphaeus cornutus (FISH.) and Nalassus convexus (COM.) are recorded from Poland, but not included in Polish fauna.

Pactola kuscheli sp. nov. (Coleoptera: Curculionidae), a potential cause for the decline of the threatened New Caledonian conifer, Agathis montana de Laubenfels, 1969 (Araucariaceae)

Agathis montana is a conifer endemic to New Caledonia and one of the most threatened (Critically Endangered) plant species of this area. The exact causes of its decline are still unknown. The authors describe the characteristics of A. montana, the history and methodology of population monitoring, and possible factors causing the decline and habitat degradation. In 2014, some twigs showing symptoms suggesting attack by wood‐boring insects were collected. From these, imagines of an unknown weevil species belonging to the genus Pactola Pascoe (Eugnomini) were reared. The new species is described as Pactola kuscheli Mazur sp. nov. Colour photographs of the dorsal habitus, line drawings of taxonomically relevant structures and a key to all New Caledonian species of Pactola are provided. The biology of P. kuscheli Mazur sp. nov. is described and its role as a key threatening process of A. montana is discussed.