Alexander G. Bugrov

Comparative FISH analysis of distribution ofB chromosome repetitive DNA in A an d B chromosomes in two subspecies of Podisma sapporensis (Orthoptera, Acrididae)

FISH analysis of B chromosome repetitive DNA distribution in A and B chromosomes of two subspecies of Podisma sapporensis (P. s. sapporensis and P. s. krylonensis) was performed. In the B chromosomes, C-positive regions contained homologous DNA repeats present also in some C-positive A chromosome regions. Most C-negative regions contained DNA repeats characteristic of A chromosome euchromatic regions. The two subspecies analyzed differed in the location of A chromosome regions enriched with repeats homologous to repeats of B chromosomes. The only common region enriched with these B chromosome repeats in both subspecies was the X chromosome pericentromeric region. The origin of B chromosomes in P. sapporensis is discussed.

DNA content of the B chromosomes in grasshopper Podisma kanoi Storozh. (Orthoptera, Acrididae)

A DNA library derived from the B chromosome of Podisma kanoi was obtained by chromosome microdissection. A total of 153 DNA clones were isolated from the microdissected DNA library. Twenty of them were sequenced. A comparison of B chromosome DNA sequences with sequences of other species from the DDBJ/GenBank/EMBL database (http://www.ddbj.nig.ac.jp/) was performed. Different patterns of signals were observed after FISH with labeled cloned DNA fragments. FISH signals with cloned DNA fragments painted either whole Bs or their different regions. Some clones also gave signals in pericentromeric regions of A chromosomes. Other cloned DNA fragments gave only background-like signals on A and B chromosomes. Comparative FISH analysis of B chromosomes in Podisma kanoi and P. sapporensis with DNA probes derived from the Bs of these species revealed homologous DNA that was confined within pericentromeric and telometric regions of the B chromosome in P. kanoi. In contrast to the B chromosomes in P. sapporensis containing large regions enriched with rDNA, only a small cluster of rDNA was detected in one of the examined B chromosomes in P. kanoi. The data strongly suggest an independent origin of B chromosomes in two closely related Podisma species.

Molecular phylogeny of Palaearctic genera of Gomphocerinae grasshoppers (Orthoptera, Acrididae)

Abstract.  Molecular phylogenetic methods were used to examine morphologically based hypotheses concerning the taxonomic structure and relationships of the grasshopper subfamily Gomphocerinae. Two mitochondrial gene (cytochrome b and cytochrome oxidase subunit I) sequences were determined for twenty‐five species representing eleven Palaearctic genera. The studied Gomphocerinae species constituted a monophyletic group; furthermore, the earlier division of Gomphocerinae into tribes was supported, with each tribe monophyletic. There was no support for various systems uniting Stenobothrini and Gomphocerini into one tribe. Two separate clusters were discerned in Gomphocerini and two tribes were distinguished – Gomphocerini (genera Aeropus, Stauroderus, Chorthippus) and Stenobothrini (genera Omocestus, Stenobothrus).

Chromosomal Localization of Ribosomal and Telomeric DNA Provides New Insights on the Evolution of Gomphocerinae Grasshoppers

Chromosome location of ribosomal DNA (rDNA) and telomeric repeats was analysed in mitotic chromosomes of 15 species of Gomphocerinae grasshoppers belonging to the tribes Arcypterini, Gomphocerini, Stenobothrini, and Chrysochraontini. Two types of rDNA distribution were found in the Gomphocerini tribe. Type 1, found in 9 species, was characterized by the presence of rDNA in the short arm of the long biarmed chromosomes 2 and 3 and, in some species, also in the X chromosome. Type 2 was found only in Aeropus sibiricus and Stauroderus scalaris and consisted in the presence of pericentromeric rDNA blocks in all chromosomes. A comparison of rDNA distribution in Gomphocerini species with 2n♂ = 23, 2n♂ = 21, and 2n♂ = 17 suggested the possible involvement of chromosome 6 in the ancestral karyotype (2n♂ = 23) in 1 of the 3 centric fusions that decreased the chromosome number in these species. In the tribe Stenobothrini, Stenobothrus eurasius carried a single rDNA cluster in the X chromosome, likewise 2 Spanish species previously analysed, but Omocestus viridulus unusually showed a single rDNA cluster in the longest autosome. Telomeric repeats were located primarily on the ends of chromosome arms. In 2 species, however, we observed the presence of interstitial clusters outside telomeric regions. The first one, Aeropus sibiricus, exhibited a polymorphic interstitial site of telomeric repeats in chromosome 6 as a consequence of a paracentric inversion. Most remarkably, Chorthippus jacobsoni showed the presence of telomeric repeats in the pericentric regions of the 3 biarmed chromosome pairs originated by centric fusion, thus suggesting that these rearrangements were not of the Robertsonian type but true centric fusion with a probable generation of dicentric chromosomes.

Karyotype evolution and chromosome C-banding patterns in some Podismini grasshoppers (Orthoptera, Acrididae)

SUMMARYThe karyotype and C-band patterns in the spermatogenesis of some species of the Podismini grasshopper are reported. The paracentromeric, interstitial, and telomeric C-bands in 14 species were observed. On the basis of comparative analysis of C-band patterns in 23- and 21-chromosome groups the hypothesis of tandem translocation of small or small and medium chromosomes in the karyotypic evolution of Podismini is suggested.

Origin of Chromosomal Rearrangement: Phylogenetic Relationship Between X0/XX and XY/XX Chromosomal Races in the Brachypterous Grasshopper Podisma sapporensis (Orthoptera: Acrididae)

Abstract The brachypterous grasshopper Podisma sapporensis Shiraki (Orthoptera: Acrididae) consists of two major chromosomal races with different sex chromosome systems. In the X0/XX race, the diploid number of chromosomes is 2n = 23 (X0) in males and 2n = 24 (XX) in females. In the XY/XX race, the diploid number is 2n = 22 (XY) in males and 2n = 22 (XX) in females, owing to Robertsonian fusion between an autosome and the X chromosome. The X0/XX and XY/XX races are allopatrically distributed, and each race contains geographical populations characterized by different chromosomal inversions. A previous hypothesis suggested that the XY/XX race was derived from the X0/XX race. To test this hypothesis, we examined mitochondrial DNA (mtDNA) sequence variation in two regions (cytochrome oxidase subunit II and 16S ribosomal DNA) among 29 P. sapporensis individuals representing five X0/XX and 12 XY/XX populations. The maximum parsimony tree of mtDNA indicated that neither of the two chromosomal races was monophyletic. Northern populations of XY/XX race did not join a clade consisting of more southern XY/XX populations but were included in a clade consisting of X0/XX populations. On the basis of these results, we propose two hypotheses for the differentiation between the northern and southern XY/XX populations. First, the XY/XX karyotype may have occurred more than once. Second, introgression of mtDNA may have occurred across adjacent populations possessing different karyotypes.

Neo-XY chromosome sex determination in four species of the pamphagid grasshoppers (Orthoptera, Acridoidea, Pamphagidae) from Bulgaria

Summary The karyotypes and C-banding patterns in the spermatogenesis of the Pamphagidae grasshoppers Paranocaracris bulgaricus (Ebn. et Drenow), Paranocarodes straubei (Fieb.), Paranocarodes chopardi Peshev and Asiotmethis limbatus (Charp.) from Bulgaria with 2n = 18 (16AA + neo-XY) are reported. The evolution of the neoXY/neo-XX sex determination in Pamphagidae is discussed.

C-heterochromatin in chromosomes of Ophiogomphus cecilia cecilia (Four.) (Anisoptera: Gomphidae) with notes on the sex chromosome origin in the species

Abstract The karyotype of Ophiogomphus cecilia cecilia (2n_=23, X0) was analyzed using C-banding technique. All autosomes possess terminal C-bands. The Xchromosome is the largest element of the set and it consists of heterochromatic region at one of the ends and euchromatic part with three interstitial heterochromatic blocks. Possible ways of the formation of the X-chromosome are considered.

Molecular cytogenetic analysis reveals the existence of two independent neo-XY sex chromosome systems in Anatolian Pamphagidae grasshoppers

BackgroundNeo-XY sex chromosome determination is a rare event in short horned grasshoppers, but it appears with unusual frequency in the Pamphagidae family. The neo-Y chromosomes found in several species appear to have undergone heterochromatinization and degradation, but this subject needs to be analyzed in other Pamphagidae species. We perform here karyotyping and molecular cytogenetic analyses in 12 Pamphagidae species from the center of biodiversity of this group in the previously-unstudied Anatolian plateau.ResultsThe basal karyotype for the Pamphagidae family, consisting of 18 acrocentric autosomes and an acrocentric X chromosome (2n♂ = 19, X0; 2n♀ = 20, XX), was found only in G. adaliae. The karyotype of all other studied species consisted of 16 acrocentric autosomes and a neo-XY sex chromosome system (2n♂♀ = 18, neo-XX♀/neo-XY♂). Two different types of neo-Y chromosomes were found. One of them was typical for three species of the Glyphotmethis genus, and showed a neo-Y chromosome being similar in size to the XR arm of the neo-X, with the addition of two small subproximal interstitial C-blocks. The second type of the neo-Y chromosome was smaller and more heterochromatinized than the XR arm, and was typical for all Nocarodeini species studied. The chromosome distribution of C-positive regions and clusters of ribosomal DNA (rDNA) and telomeric repeats yielded additional information on evolution of these neo-XY systems.ConclusionMost Pamphagidae species in the Anatolian region were found to have neo-XY sex chromosome systems, belonging to two different evolutionary lineages, marked by independent X-autosome fusion events occurred within the Trinchinae and Pamphaginae subfamilies. The high density of species carrying neo-XY systems in the Anatolian region, and the different evolutionary stage for the two lineages found, one being older than the other, indicates that this region has a long history of neo-XY sex chromosome formation.

Origin and Evolution of the Neo-Sex Chromosomes in Pamphagidae Grasshoppers through Chromosome Fusion and Following Heteromorphization

In most phylogenetic lineages, the evolution of sex chromosomes is accompanied by their heteromorphization and degradation of one of them. The neo-sex chromosomes are useful model for studying early stages of these processes. Recently two lineages of the neo-sex chromosomes on different stages of heteromorphization was discovered in Pamphagidae family. The neo-sex chromosome heteromorphization was analyzed by generation of DNA probes derived from the neo-Xs and neo-Ys followed with chromosome painting in nineteen species of Pamphagidae family. The homologous regions of the neo-sex chromosomes were determined in closely related species with the painting procedure and image analysis with application of the Visualization of the Specific Signal in Silico software package. Results of these analyses and distribution of C-positive regions in the neo-sex chromosomes revealed details of the heteromorphization of the neo-sex chromosomes in species from both phylogenetic lineages of Pamphagidae grasshoppers. The hypothetical mechanism of the neo-Y degradation was suggested. It includes expansion of different repeats from the proximal neo-Y chromosome region by inversions, spreading them towards distal region. Amplification of these repeats leads to formation of C-positive regions and elimination of the C-negative regions located between them.