Molecular and classical chromosomal techniques reveal diversity in bushcricket genera of Barbitistini (Orthoptera).

The cytogenetic characteristics of 17 species of bushcricket belonging to eight genera of the tribe Barbitistini were examined by fluorescence in situ hybridization with 18S rDNA and (TTAGGn) telomeric as probes and by C-banding, silver, and fluorochrome staining. These markers were used to understand chromosomal organization and evolutionary relationships between genera or species within the same genus. The number of 18S rDNA clusters per haploid genome that co-localized with active nucleolus organizer regions (NORs) ranged from one to five, with the most common pattern being the presence of one NOR-bearing chromosome. This ribosomal cistron was preferentially located in the paracentromeric region of autosomes and very rarely in the sex chromosome. The results demonstrated coincidence between the localization of major ribosomal genes and active NORs and the position of C-band and GC-rich regions. The rDNA/NOR distribution and the composition of chromosome heterochromatin proved to be good cytogenetic markers for distinguishing species and phylogenetic lines and for understanding the genomic differentiation and evolution of Barbitistini. A comparison of cytogenetic and morphological or behavioral traits suggests that morphological and behavioral specialization in this group was not followed by major karyotype modification (except for Leptophyes). However, the occurrence and distribution of different repetitive DNA sites tends to vary among the taxa.

Evidence for two karyotypic variants of the lesser horseshoe bat ( Rhinolophus hipposideros , Chiroptera, Mammalia) in Central Europe.

Three different diploid chromosome numbers (2n = 54, 56 and 58) have been reported in the lesser horseshoe bat, Rhinolophus hipposideros. Asia Minor and the Middle East are inhabited by R. hipposideros specimens with 58 chromosomes. In Europe, specimens with 56 chromosomes have been recorded from several localities in the Czech Republic, Slovakia, Italy and Greece. Up to now, specimens with 54 chromosomes have been reported only from Spain and possibly from Switzerland. With the record of 54 chromosomes in specimens from Germany presented here, the distributional area of this variant is expanded into Central Europe. According to the cytogenetic data presently available, we presume that the European R. hipposideros population is divided into a western form (from Spain to Germany) with a 2n = 54 karyotype and an eastern form (from the Czech Republic to Greece) with a 2n = 56 karyotype. This study presents banded karyotypes for the 2n = 54 and 2n = 56 variants for the first time. In addition, chromosomal arm homology to the vespertilionid bat species Myotis myotis revealed by chromosome painting is reported. Whether the variants could represent separate species is also discussed.

The evolution of sex differences in mate searching when females benefit: new theory and a comparative test.

Sexual selection is thought to have led to searching as a profitable, but risky way of males obtaining mates. While there is great variation in which sex searches, previous theory has not considered search evolution when both males and females benefit from multiple mating. We present new theory and link it with data to bridge this gap. Two different search protocols exist between species in the bush-cricket genus Poecilimon (Orthoptera): females search for calling males, or males search for calling females. Poecilimon males also transfer a costly nuptial food gift to their mates during mating. We relate variations in searching protocols to variation in nuptial gift size among 32 Poecilimon taxa. As predicted, taxa where females search produce significantly larger nuptial gifts than those where males search. Our model and results show that search roles can reverse when multiple mating brings about sufficiently strong material benefits to females.

Chromosome number reduction accompanied by extensive heterochromatin addition in the bat Glauconycteris beatrix (Mammalia; Chiroptera, Vespertilionidae).

The presumed ancestral karyotype of the bat family Vespertilionidae consists of 44 chromosomes with a fundamental number of autosomal arms (FNa) of 50. Previously, only two of the roughly 350 vespertilionid species have been reported with 2n lower than 26. In this paper we report the 2n = 22 karyotype of the African vespertilionid Glauconycteris beatrix which shows an X-autosome translocation and extended, paracentromeric, chromomycin-A3-positive heterochromatin.

A comparative ZOO-FISH analysis in bats elucidates the phylogenetic relationships between Megachiroptera and five microchiropteran families.

Fluorescence in-situ hybridization with human whole chromosome painting probes (WCPs) was applied to compare the karyotypes of members of five bat families. Twenty-five evolutionarily conserved units (ECUs) were identified by ZOO-FISH analysis. In 10 of these 25 ECUs, thorough GTG-band comparison revealed an identical banding pattern in all families studied. Differences in the remaining ECUs were used as characters to judge the phylogenetic relationships within Chiroptera. Close relationships were found between Rhinolophidae and Hipposideridae. Also closely related are the representatives of the yangochiropteran families Phyllostomidae (genus studied: Glossophaga, Volleth et al. 1999), Molossidae and Vespertilionidae. All microchiropteran species studied here share four common features not found in the megachiropteran species Eonycteris spelaea. Two of these are considered as derived characters with a high probability of parallel evolution. On the other hand, Eonycteris shares one common, probably derived feature with the rhinolophoid families Rhinolophidae and Hipposideridae and an additional one only with Hipposideridae. At the moment, the relationships between Yangochiroptera, Rhinolophoidea and Megachiroptera must be left in an unsolved trichotomy. Comparison of neighboring segment combinations found in Chiroptera with those found in other mammalian taxa revealed six synapomorphic features for Chiroptera. Therefore, for karyological reasons, monophyly of Chiroptera is strongly supported.

Cryptic mammalian species: a new species of whiskered bat (Myotis alcathoe n. sp.) in Europe.

The analysis of morphological, behavioural and genetic characters of whiskered bats revealed a new European bat species within the family Vespertilionidae. We describe the morphology, karyology, genetic similarity, ecology and distribution of Myotis alcathoe n. sp. It closely resembles Myotis mystacinus, Myotis brandtii and Myotis ikonnikovi in morphology, but all four species show clear genetic differences in two mitochondrial genes (ND1 and 12S rRNA). Myotis alcathoe n. sp. is the smallest species among the European whiskered bats and uses the highest-frequency echolocation calls of all the European Myotis species. It prefers to hunt in small valleys with deciduous trees and flowing water, which is an endangered habitat. Records from Greece and Hungary indicate a distribution range in south-eastern Europe.

Karyotype comparison and phylogenetic relationships of Pipistrellus-like bats (Vespertilionidae; Chiroptera; Mammalia).

Detailed karyotype descriptions of 20 Pipistrellus-like bat species belonging to the family Vespertilionidae are presented. For the first time, chromosomal complements of four species, i.e. Pipistrellus stenopterus (2n = 32), P. javanicus (2n = 34), Hypsugo eisentrauti (2n = 42) and H. crassulus (2n = 30) are reported. A Pipistrellus kuhlii-like species from Madagascar represents a separate species distinguished from the European Pipistrellus kuhlii (2n = 44) by a diploid chromosome number of 42. Banded karyotypes are presented for the first time for Scotozous dormeri, Hypsugo capensis, Hesperoptenus blanfordi, Tylonycteris pachypus and robustula. Chromosomal evolution in the family Vespertilionidae is characterized by the conservation of entire chromosomal arms and reductions in diploid chromosome number via Robertsonian fusions. Less frequently, centric fissions, para- and pericentric inversions and centromere shifts were found to have occurred. In several cases a certain type of chromosomal change predominates in a karyotype. Examples of this are the acquisition of interstitial heterochromatic bands in Tylonycteris robustula, and centric shifts in P. javanicus, H. eisentrauti and Hesp. blanfordi. The species examined here belong to three tribes, i.e. Pipistrellini, Vespertilionini and Eptesicini, which are distinguished by chromosomal characteristics. According to our results, the species Pipistrellus (Neoromicia) capensis belongs to the Vespertilionini and not to the Pipistrellini. We therefore propose to elevate the subgenus Neoromicia to generic rank.

Carotenoids in the spermatophores of bushcrickets (Orthoptera: Ephippigerinae).

During mating male bushcrickets transfer large spermatophores, which have been demonstrated to play an important role in female nutrition and egg production. Until now only relatively unspecific substances such as water and proteins were known to be present within these spermatophores. We found that in the bushcricket Ephippiger zelleri the spermatophores contain substantial amounts of carotenoids (mainly lutein and zeaxanthin) that are also found in the eggs of this species. Carotenoids are well known for their positive effects on survival and reproduction in animals. This is the first example, to our knowledge, where such specific vitamin-like substances were found to be transferred from male to female during mating.

Chromosome evolution in the genus Poecilimon (Orthoptera, Tettigonioidea, Phaneropteridae).

Cytotaxonomic analysis of 20 species and subspecies of the genus Poecilimon using C-banding pattern, chiasma frequency, and morphometric characteristics of the chromosomes were described. Using a cladistic analysis the chromosome data provided a basis to produce a phylogenetic tree which was compared with a tree based on morphological characters and DNA sequence data. There are important differences in the grouping of data sets to species obtained on the basis of morphology/DNA analyses and that based on chromosomes. The explanation of the differences between C-banding patterns and taxonomic proximity is probably that the C-banding pattern changes quickly as the result of the high degree of variation of constitutive heterochromatin.

The chemical composition of the spermatophore in some species of phaneropterid bushcrickets (Orthoptera: Tettigonioidea).

Male bushcrickets transfer large spermatophores of up to 40% of male body mass to the females during mating. These large nuptial gifts are later consumed by the female and have been shown to affect the size and number of eggs laid after mating in some species. The composition of the spermatophores in five species of phytophagous phaneropterid bushcrickets (genera Ancistrura, Barbitistes, Metaplastes, Poecilimon) was examined in respect to water content, elemental composition, protein concentration, and the content of lipids, carbohydrates, urea and uric acid. In addition, the amino acid composition of the spermatophore was compared with that of the egg proteins. In all species, water content was found to be about 85% of wet mass. Eleven to 16% of the dry mass consisted of nitrogen, corresponding to a protein content of about 70 to more than 90% of the dry mass. Urea and uric acid were only present in traces. The proteins contained a high amount of glycine (about 26mol %), together with asparagine/aspartic acid, 12% and glutamic acid, 11%, which differed distinctly from the amino acid composition of the egg proteins. The results are discussed with respect to the advantages the male may receive by producing these large nuptial gifts.

A new biophysical method to determine the gain of the acoustic trachea in bushcrickets.

A method is described for measuring the gain (i.e., the change of amplitude and phase angle) for sounds that propagate to the internal surface of the tympana in ears working as pressure difference receivers. The gain of the acoustic trachea has been measured in two similarly sized and closely related species of bushcrickets, in which the acoustic spiracles and tracheae differ markedly in size. The amplitude part of the gain is much larger in the species with the larger acoustic spiracle, whereas the phase part is very similar in the two species. The method is compared with other methods, which in the past have been used for estimating the gain of sound pathways inside animal bodies.