Eight species belonging to five true bug families were analyzed using DAPI/CMA3-staining and fluorescence in situ hybridization (FISH) with telomeric (TTAGG)n and 18S rDNA probes. Standard chromosomal complements are reported for the first time for Deraeocoris rutilus (Herrich-Schäffer, 1838) (2n=30+2m+XY) and Deraeocoris ruber(Linnaeus, 1758) (2n=30+2m+XY) from the family Miridae. Using FISH, the location of a 18S rDNA cluster was detected in these species and in five more species: Megaloceroea recticornis (Geoffroy, 1785) (2n=30+XY) from the Miridae; Oxycarenus lavaterae (Fabricius, 1787) (2n=14+2m+XY) from the Lygaeidae s.l.; Pyrrhocoris apterus (Linnaeus, 1758) (2n=22+X) from the Pyrrhocoridae; Eurydema oleracea (Linnaeus, 1758) (2n=12+XY) and Graphosoma lineatum (Linnaeus, 1758) (2n=12+XY) from the Pentatomidae. The species were found to differ with respect to location of a 18S rRNA gene cluster which resides on autosomes in Oxycarenus lavaterae and Pyrrhocoris apterus, whereas it locates on sex chromosomes in other five species. The 18S rDNA location provides the first physical landmark of the genomes of the species studied. The insect consensus telomeric pentanucleotide (TTAGG)n was demonstrated to be absent in all the species studied in this respect, Deraeocoris rutilus, Megaloceroea recticornis, Cimex lectularius Linnaeus, 1758 (Cimicidae), Eurydema oleracea, and Graphosoma lineatum, supporting the hypothesis that this motif was lost in early evolution of the Heteroptera and secondarily replaced with another motif (yet unknown) or the alternative telomerase-independent mechanisms of telomere maintenance. Dot-blot hybridization analysis of the genomic DNA from Cimex lectularius, Nabis sp. and Oxycarenus lavaterae with (TTAGG)n and six other telomeric probes likewise provided a negative result.

Abstract. Bugs (Insecta: Heteroptera) are frequently used as examples of unusual cytogenetic characters, and the family Cimicidae is one of most interest in this respect. We have performed a cytogenetic study of the common bed bug Cimex lectularius Linnaeus, 1758 using both classical (Schiff-Giemsa and AgNO 3 -staining) and molecular cytogenetic techniques (base-specifi c DAPI/CMA 3 fl uorochromes and FISH with an 18S rDNA probe). Males originated from a wild population of C. lectularius were found to have 2n = 26 + X 1 X 2 Y, holokinetic chromosomes, 18S rRNA genes located on the X 1 and Y chromosomes; achiasmate male meiosis of a collochore type; MI and MII plates nonradial and radial respectively.

Genomes of numerous diploid plant and animal species possess traces of interspecific crosses, and many researches consider them as support for homoploid hybrid speciation (HHS), a process by which a new reproductively isolated species arises through hybridization and combination of parts of the parental genomes, but without an increase in ploidy. However, convincing evidence for a creative role of hybridization in the origin of reproductive isolation between hybrid and parental forms is extremely limited. Here, through studying Agrodiaetus butterflies, we provide proof of a previously unknown mode of HHS based on the formation of post-zygotic reproductive isolation via hybridization of chromosomally divergent parental species and subsequent fixation of a novel combination of chromosome fusions/fissions in hybrid descendants. We show that meiotic segregation, operating in the hybrid lineage, resulted in the formation of a new diploid genome, drastically rearranged in terms of chromosome number. We also demonstrate that during the heterozygous stage of the hybrid species formation, recombination was limited between rearranged chromosomes of different parental origin, representing evidence that the reproductive isolation was a direct consequence of hybridization.

The phylogenies of all eight European species of Philaenus were estimated from cytochrome oxidase subunit I, cytochrome B and internal transcribed spacer 2 (ITS2) fragments of DNA using phylogenetic reconstruction methods: maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI) analyses. Based on the topologies of all obtained phylogenetic trees, the monophyly of Philaenus is well supported, being congruent with morphological, ecological and chromosomal data. Three phylogenetic lineages were distinguished in the mitochondrial and combined (mtDNA with ITS2) trees. The first lineage is represented by only one species, Philaenus maghresignus, which inhabits Maghreb and southern Spain. Clade A includes three species: P. tarifa (Southern Iberia), P. italosignus (Sicily and Southern Italy) and P. signatus (the Balkans and Middle East). In clade B two subclades were recognized: B1 represented by P. loukasi (Southern Balkans) and P. arslani (Middle East), and B2 comprising P. spumarus (the most widespread Palaearctic species) and P. tesselatus (from Southern Iberia and Maghreb). These clades were also retrieved in trees reconstructed from nuclear sequences. However, four species (P. maghresignus, P. tarifa, P. italosignus and P. signatus) showed unresolved polytomy at the base of the nuclear tree. Clade A together with P. maghresignus clustered with the 'signatus' group defined from morphology, and clade B with the 'spumarius' group; these might be considered separate subgenera. Genetic distances in mitochondrial DNA between ingroup species ranged from 14.0% between P. signatus and P. spumarius to 2.4% between P. tesselatus and P. spumarius. By contrast, genetic divergence of ITS2 between ingroup species was very low, at most 2.1%. The divergence of Philaenus species is estimated to have occcurred between 7.9 and 0.6 Ma. Possibly three main speciation events occurred: the first at the Miocene/Pliocene boundary (c. 5.5 Ma) for deeper splits; the second between 4.2 and 2.5 Ma in the Pliocene, when pairs of more closely related species diverged; and the most recent during the Pleistocene glaciations, when the separation of P. tesselatus and P. spumarius took place. The species status of all Philaenus species is confirmed except for P. tesselatus.

Male karyotypes of Elasmotropis testacea (Herrich-Schaeffer, 1835), Tingis cardui (Linnaeus, 1758), Tingis crispata (Herrich-Schaeffer, 1838), and Agramma femorale Thomson, 1871 (Heteroptera, Cimicomorpha, Tingidae) were analyzed using conventional chromosome staining and FISH with 18S rDNA and (TTAGG)n telomeric probes. The FISH technique was applied for the first time in the Tingidae. In spite of the fact that all species showed the same chromosome number (2n = 12 + XY), they have significant differences in the number and position of rDNA loci. FISH with the classical insect (TTAGG)n probe produced no signals on chromosomes suggesting telomeres in lace bugs to be of some other molecular composition. Tingidae share absence of the (TTAGG)n telomeric sequence with all so far studied taxa of the advanced true bug infraorders Cimicomorpha and Pentatomomorpha.

Male Nabis (Aspilaspis ) indicus (Stål), N. (A .) viridulus Spinola, Himacerus (Himacerus ) mirmicoides (O. Costa) (2n 0/32'/ XY) and Prostemma guttula (Fabricius) (2n0/26'/XY) were studied using C-banding, silver nitrate staining and basespecific fluorochrome (DAPI and CMA 3 ) staining. N. indicus differed from N. viridulus in distribution pattern of C-bands, which were telomeric in the former while interstitial in the latter. H. mirmicoides showed interstitial C-bands in the majority of autosomes. P. guttula had no conspicuous C-bands in other chromosomes, but only in the Y, which was totally heterochromatic. C-heterochromatin was labelled with DAPI, indicating that it was AT-rich. In every species, both X and Y chromosomes were NOR-bearing, and the NOR regions were GC-rich. In H. mirmicoides and P. guttula, NORs showed sub-median location in the X and distal in the Y, such a pattern being probably common in Nabidae. The present paper provides new information on the genome organization and new cytological markers useful for a better insight into karyotype evolution of nabid species.

No abstract

Karyotypes of six species belonging to three main clades of parasitoid Hymenoptera, the superfamilies Ichneumonoidea (Ichneumonidae: Ichneumon amphibolus), Cynipoidea (Cynipidae: Diplolepis rosae) and Chalcidoidea (Eurytomidae: Eurytoma robusta, Eu. serratulae and Eu. compressa, and Torymidae: Torymus bedeguaris) were studied using FISH with 18S rDNA and telomeric (TTAGG)n probes. Haploid karyotypes of D. rosae, Eu. robusta and Eu. serratulae carried the only 18S rDNA hybridization signal, whereas those of I. amphibolus and Eu. compressa carried three and two rDNA clusters respectively. In addition, three rDNA sites were visualized in the aneuploid female of T. bedeguaris. The number of rDNA clusters in parasitoid Hymenoptera generally correlates to the chromosome number. Apart from the overwhelming majority of the studied species of aculeate Hymenoptera, no hybridization signals were obtained from FISH with the telomeric (TTAGG)n probe in the examined parasitoid species. These data suggest absence of the canonical (TTAGG)n insect telomeric motif in the Ichneumonoidea, Cynipoidea and Chalcidoidea, and perhaps in parasitoid Hymenoptera in general.