Chromosomes of the males of five species of Odontura, belonging to the subgenera Odontura and Odonturella, were analyzed. Intensive evolution of the karyotype was recorded, both in terms of changes in the numbers of chromosomes (from 2n = 31 to 27) and the sex chromosome system (from X0 to neo-XY and X0 to neo-X1X2Y). Karyotype evolution was accompanied by tandem autosome fusions and interspecific autosomal and sex chromosome differentiation involving changes in the locations of nucleolar organizer regions, NORs, which were revealed by silver impregnation and confirmed by FISH using an 18S rDNA probe. O. (Odonturella) aspericauda is a polytypic species with X0 and neo-X1X2Y sex determination. The latter system is not common in tettigoniids. It possibly originated by a translocation of a distal segment of the original X chromosome onto a medium sized autosome, resulting in a shortened neo-X1 and a metacentric neo-Y. The remaining autosome homologue became the neo-X2 chromosome. This shift from X0 to neo-X1X2Y is supported by the length of the X chromosome and location of the NOR/rDNA. and Elżbieta Warchałowska-Śliwa, Anna Maryańska-Nadachowska, Beata Grzywacz, Tatjana Karamysheva, Arne W. Lehmann, Gerlind U.C. Lehmann, Klaus-Gerhard Heller.
We report the karyotype characteristics including chromosome numbers of Saga campbelli campbelli, S. c. gracilis, and S. rammei using the following classical cytogenetic methods: C-banding, silver staining, and fluorochrome staining DAPI and CMA3. We also present FISH data showing the distribution of telomeric repeats and 18S rDNA on the chromosomes of these species and the results of similar studies cited in the literature on S. hellenica, S. natoliae, and S. rhodiensis. The five European Saga species exhibit a high rate of karyotype evolution. In addition to changes in chromosome number and morphology (by chromosomal inversion and/or chromosome fusion), interspecific autosomal differentiation involved changes in the distribution and quantity of constitutive heterochromatin and GC-rich regions, as well as the number and location of NORs. In the present study we focused on testing a hypothetical model of karyotype evolution in Saga, with particular reference to the cytogenetic mapping of rDNA and telomeric sequences. Variation in the distribution of rDNA and location of Ag-NORs are novel phylogenetic markers for the genus Saga.
A cytogenetic investigation was performed in eight species of the spittlebug genus Philaenus using silver-NOR (AgNOR)-banding and fluorescence in situ hybridization (FISH) with 18S rDNA and (TTAGG)n telomeric probes. This is the first application of FISH technique in the Auchenorrhyncha, a suborder of the Hemiptera. FISH along with the rDNA probe revealed differences between species in the number and chromosomal location of major ribosomal RNA gene sites, the so-called nucleolar organizer regions (NORs). However, we found a lack of perfect correlation between the results of AgNOR-staining and rDNA-FISH in the detection of NORs. FISH with the telomeric probe confirmed that the chromosome ends of the Philaenus species are composed of the (TTAGG)n nucleotide sequence, which is a common motif of insect telomeres., Anna Maryanska-Nadachowska, Valetnina G. Kuznetsova, Tatyana V. Karamysheva., and Obsahuje seznam literatury
The present study focused on the evolution of the karyotype in 21 taxa of the genus Isophya, which was done by mapping the location on the chromosomes of ribosomal RNA (rRNA) coding genes using fluorescence in situ hybridization (FISH) with an 18S rDNA probe and using silver staining (AgNO3) to evaluate the activity of major rDNA clusters. Since the chromosome number and sex determination do not vary in this genus, the above markers were used in a detailed comparison of the cytogenetic features of species of Isophya. The species analyzed were placed into three groups based on the location of rDNA on their chromosomes: (1) rDNA-FISH signals only on the two long pairs of autosomes, (2) rDNA-FISH signals on one long and one short pair of autosomes, and (3) rDNA-FISH signals on three to five different sized pairs of autosomes. These groupings partly correspond to the morphological groupings proposed in earlier studies. One long pair of autosomes frequently carried rDNA in all the Isophya species and probably is a plesiomorphic character for these taxa. The cytogenetic mapping revealed great variability among Isophya species in the chromosomal location of major rDNA clusters. Our results suggest that the observed variation in the number of rDNA clusters can be an important species-group specific phylogenetic marker. Analysis of 18S rDNA hybridization signals showed that the evolutionary dynamics of rDNA in this genus is remarkably high and accompanied by changes in the structure of chromosomes bearing rDNA at an inter- and intra-specific level. The telomeric sequence (TTAGG)n hybridized with the termini of most of chromosomes, however, some chromosome ends lacked signals probably due to a low copy number of telomeric repeats. and Beata Grzywacz, Anna Maryańska-Nadachowska, Dragan P. Chobanov, Tatjana Karamysheva, Elżbieta Warchałowska-Śliwa.
Diachasmimorpha longicaudata (Hymenoptera: Braconidae) is a parasitoid wasp widely used in the biological control of fruit flies. In this paper, we present a detailed analysis of the karyotype of this species based on the results of classical and molecular cytogenetic techniques. The cytogenetic analysis confirmed the male and female chromosome numbers previously reported (n = 20, 2n = 40). The entire short arm of most chromosomes is made up of a large constitutive heterochromatic segment. The high heterochromatin content differentiates D. longicaudata from other braconid species. Fluorescence in situ hybridization (FISH) using autologous 18S rDNA probes revealed six clusters of rDNA, i.e. six nucleolar organizer regions (NORs), in the heterochromatic short arms of different chromosomes in the haploid male karyotype. This number is exceptionally high for Hymenoptera, which usually have two NORs in the diploid complement. It is noteworthy that these rDNA-FISH experiments represent the first use of this technique on a braconid species using autologous probes. Since Ag-NOR-bands were coincident with C-positive bands on metaphase chromosomes, it was not possible to identify active nucleoli. The physical characteristics of the D. longicaudata karyotype, especially the content and distribution of heterochromatin and the number and location of rDNA clusters, contribute to a better understanding of the structure and organization of braconid chromosomes and provide a basis for genomic and evolutionary studies., Leonela Carabajal Paladino ... [et al.]., and Obsahuje seznam literatury
1_Chromosomes of six European species (one with two subspecies) of Orthoptera belonging to the tribes Ephippigerini and Bradyporini were analyzed using C-banding, Ag-NOR, DAPI (AT-rich)/CMA3 (GC-rich) staining and fluorescence in situ hybridization (FISH) using the 18S rDNA and (TTAGG)n telomeric probes with the aim to better understand chromosomal organization and evolutionary relationships between genera and subgenera within and across both tribes. The evolution of karyotypes was studied in terms of changes in chromosome number (2n) and morphology (FN, the fundamental number – i.e. the number of chromosome arms including the X chromosome). The ancestral 2n = 31 was reduced to 2n = 29 (FN = 31) and 27 (FN = 31) by one or two Robertsonian fusions in the Ephippigerini. Whereas in the Bradyporini 2n = 27 (FN = 32) as a result of two Robertsonian translocations and a pericentric inversion in the X chromosome. The quantity of heterochromatin in GC-rich regions distinguished the karyotypes of Ephippigerini (only a single CG-rich band on one autosome pair) from those of Bradyporini (CG-rich bands on all chromosomes). FISH using the 18S rDNA probe localized 1–3 rDNA clusters to autosomes and/or to the X chromosome in all species examined. The rDNA loci coincided with active NORs as determined by Ag-NOR staining. A comparison of the location of the single NOR/rDNA in two species of the genus Steropleurus (Ephippigerini) suggests that the reduced chromosome number in S. pseudolus results from a Robertsonian fusion between two pairs of autosomes, one of them carrying the NOR/rDNA as in S. stalii (and also in E. ephippiger)., 2_Whereas the karyotypes of three species of the genus Bradyporus, though showing the same chromosome number and morphology, differed in the number and distribution of NORs/rDNA sites [one autosomal in B. (B.) dasypus versus three in B. macrogaster and B. (C.) oniscus, two of them X-linked]. Trends in karyotype diversification of the taxa based on the present data and previous research are discussed. In some individuals belonging to the species Bradyporus (B.) dasypus and B. (C.) m. macrogaster B chromosomes (Bs) were detected: acrocentric (the smallest elements in the complement) and submetacentric (similar to medium-sized autosomes), respectively., Elzbieta Warchalowska-Sliwa ... [et al.]., and Obsahuje seznam literatury
Myeloproliferativní neoplázie (MPN) představují skupinu klonálních onemocnění hematopoetické kmenové buňky s fenotypickými změnami, které jsou důsledkem zvýšené myeloidní proliferace se zachovanou maturací a akumulací myeloidních buněk v periferní krvi: leukocytózou, trombocytózou a zvýšeným počtem červených krvinek nebo různou kombinací proliferace jednotlivých buněčných linií. Cytogenetické analýzy odhalily opakující se chromozomové změny u 5–45 % nemocných, ale specifická chromozomová změna zatím nebyla nalezena. K častým chromozomovým změnám, které u MPN nacházíme, patří především ztráty genetického materiálu v podobě delecí dlouhých ramen chromozomu 20 (20q-), dlouhých ramen chromozomu 13 (13q-) a delece krátkých ramen chromozomu 12 (12p-). Zmnožení genetického materiálu se týká především trisomií chromozomů 8 (+8) a 9 (+9) a parciálních duplikací dlouhých ramen chromozomu 1 (1q+). K dalším pozorovaným změnám patří balancované translokace, například chromozomu 8 s různými chromozomovými partnery. Takový nález je označován jako syndrom 8p11 (EMS – eight myeloproliferative syndrome) a vždy zahrnuje gen FGFR1. Cytogenetika doplněná molekulárně cytogenetickými a genetickými metodami doplňuje diagnostiku MPN, ale má význam i pro stanovení prognózy. Určení přestavby FIP1L1/ PDGFRA u nemocných s MPN s eosinofílií umožňuje zahájit efektivní cílenou léčbu imatinibem. Přesné určení genetických změn umožňuje porozumět molekulární etiopatogenezi MPN a přispívá ke zpřesnění klasifikace, prognózy a personalizace léčby nemocných., Myeloproliferative neoplasias (MPNs) are a group of hematopoietic stem cell clonal diseases with phenotypic changes resulting from increased myeloid proliferation with preserved maturation and accumulation of myeloid cells in peripheral blood: leukocytosis, thrombocytosis and increased red blood cell count or their various combinations. Cytogenetic analyses revealed recurrent chromosomal aberrations in 5–45 % of patients, but so far, a specific chromosomal change has not been found. Frequent chromosomal aberrations observed in MNPs mainly include losses of genetic material such as deletions of the long arms of chromosome 20 (20q-), long arms of chromosome 13 (13q-) and deletions of the short arms of chromosome 12 (12p-). Gains of genetic material are particularly in the form of trisomies of chromosomes 8 (+8) and 9 (+9) and partial duplications of the long arms of chromosome 1 (1q+). Other observed changes are balanced translocations, such as those of chromosome 8 with various chromosomal partners. These findings are referred to as the 8p11 myeloproliferative syndrome (EMS), always involving the FGFR1 gene. Cytogenetic analyses supplemented with molecular cytogenetic and genetic methods play supplementary role in the diagnosis of MPNs but are also contributory prognostic determinants. Recognition of the FIP1L1/PDGFRA rearrangement in patients with NPM with eosinophilia enables initiation of targeted therapy with imatinib, significantly improving their prognosis. Accurate determination of genetic aberrations enables understanding of the molecular etiopathogenesis of MPNs and contributes to more accurate classification and more individual therapy of patients., Marie Jarošová, Milena Holzerová, Radka Nedomová, Pavla Mičková, Antonín Hluší, Karel Indrák, and Literatura 80
Fluorescence in situ hybridization (FISH) is a technique used to determine the chromosomal position of DNA and RNA probes. The present study contributes to knowledge on jumping plant-lice genomes by using FISH with 18S rDNA and telomeric (TTAGG)n probes on meiotic chromosomes of Psylla alni (2n = 24 + X), Cacopsylla mali (2n = 22 + neo-XY and 20 + neo-X1X2Y), C. sorbi (2n = 20 + neo-XY), Baeopelma foersteri (2n = 14 + X), and Rhinocola aceris (2n = 10 + X). This is the first study that has used FISH on the hemipteran superfamily Psylloidea. We found that the chromosomes of all studied species contain the insect-type telomere motif, (TTAGG)n. In C. mali and C. sorbi, the neo-sex chromosomes originating from autosome-sex chromosome fusions showed no interstitially located clusters of TTAGG repeats, suggesting their loss or inactivation. Similarly, no interstitial (TTAGG)n clusters were detected in an extremely large autosome pair of B. foersteri that most likely originated from a fusion of at least five ancestral chromosome pairs. Clusters of 18S rDNA were detected on the fused and second largest autosome pairs of B. foersteri and on one of the large autosome pairs of the remaining species. In C. mali and B. foersteri, the rDNA clusters were shown to coincide with the NORs as detected by the AgNOR method. Finally, we speculate, based on the obtained FISH markers, on the mechanisms of karyotype evolution of psylloid species differing in chromosome numbers and sex chromosome systems., Anna Maryańska-Nadachowska, Valentina G. Kuznetsova, Natalia V. Golub, Boris A. Anokhin., and Obsahuje bibliografii
The structure of the holocentric chromosomes of the rosy apple aphid, Dysaphis plantaginea (2n = 12), and pear-grass aphid, Melanaphis pyraria (2n = 8), was studied using C-banding, NOR, Giemsa and fluorochrome staining, and fluorescent in situ hybridization (FISH). Contrary to the equilocal distribution of heterochromatin typical of monocentric chromosomes, in both species C-banding evidenced a tendency of highly repetitive DNAs to be restricted to the X chromosomes. Silver staining and FISH, using a 28S rDNA probe, located rDNA genes on one telomere of each X chromosome, the only brightly fluorescent C-positive sites revealed by CMA3 staining, whereas all other heterochromatic C-bands were DAPI positive. Both species showed a noticeable amount of rDNA heteromorphism. Mitotic recombination is proposed as a possible mechanism responsible for the variation in size of rDNA.
The present paper reports some cytogenetic peculiarities observed in the Ag-NORs of Pamphagus ortolaniae chromosomes, the unusual behaviour of ribosomal sites after silver staining and the intense Ag-positive reaction of centromeric regions at spermatogonial metaphase and spermatocyte metaphase I and II. Moreover, a conclusive identification and localization of all the ribosomal clusters is provided by using heterologous rDNA FISH on spermatocyte chromosomes. 18S-28S rDNA mapped on a single chromosome pair and resulted multiclustered along the chromosomal body in three distinct serial regions, r1, r2 and r3. Surprisingly, these areas were scarcely (r1) or never (r2 and r3) detectable by silver impregnation. As in other Orthoptera and many groups of arthropods, FISH with the pentamer (TTAGG)n as the probe labelled the telomeres of all chromosomes.