Cytogenetic markers using single-sequence probes reveal chromosomal locations of tandemly repetitive genes in scleractinian coral Acropora pruinosa

The short and similar sized chromosomes of Acropora pose a challenge for karyotyping. Conventional methods, such as staining of heterochromatic regions, provide unclear banding patterns that hamper identification of such chromosomes. In this study, we used short single-sequence probes from tandemly repetitive 5S ribosomal RNA (rRNA) and core histone coding sequences to identify specific chromosomes of Acropora pruinosa. Both the probes produced intense signals in fluorescence in situ hybridization, which distinguished chromosome pairs. The locus of the 5S rDNA probe was on chromosome 5, whereas that of core histone probe was on chromosome 8. The sequence of the 5S rDNA probe was composed largely of U1 and U2 spliceosomal small nuclear RNA (snRNA) genes and their interspacers, flanked by short sequences of the 5S rDNA. This is the first report of a tandemly repetitive linkage of snRNA and 5S rDNA sequences in Cnidaria. Based on the constructed tentative karyogram and whole genome hybridization, the longest chromosome pair (chromosome 1) was heteromorphic. The probes also hybridized effectively with chromosomes of other Acropora species and population, revealing an additional core histone gene locus. We demonstrated the applicability of short-sequence probes as chromosomal markers with potential for use across populations and species of Acropora.

New cytogenetic data for three species of Pentatomidae (Heteroptera): Dichelops melacanthus (Dallas, 1851), Loxa viridis (Palisot de Beauvois, 1805), and Edessa collaris (Dallas, 1851)

In this paper, we present new cytogenetic data for three species of the family Pentatomidae: Dichelops melacanthus (Dallas, 1851), Loxa viridis (Palisot de Beauvois, 1805), and Edessa collaris (Dallas, 1851). All studied species presented holocentric chromosomes and inverted meiosis for the sex chromosomes. D. melacanthus has 2n = 12 (10A + XY); L. viridis showed 2n = 14 (12A + XY); and E. collaris showed 2n = 14 (12A + XY). C-banding was performed for the first time in these species and revealed terminal and interstitial heterochromatic regions on the autosomes; DAPI/CMA3 staining showed different fluorescent patterns. In all species, fluorescence in situ hybridization (FISH) with 18S rDNA probe identified signals on one autosomal bivalent, this being the first report of FISH application in the species D. melacanthus and L. viridis. The results obtained add to those already existing in the literature, enabling a better understanding of the meiotic behavior of these insects.

New cytogenetic data for three species of Pentatomidae (Heteroptera): Dichelops melacanthus (Dallas, 1851), Loxa viridis (Palisot de Beauvois, 1805), and Edessa collaris (Dallas, 1851)

In this paper, we present new cytogenetic data for three species of the family Pentatomidae: Dichelops melacanthus (Dallas, 1851), Loxa viridis (Palisot de Beauvois, 1805), and Edessa collaris (Dallas, 1851). All studied species presented holocentric chromosomes and inverted meiosis for the sex chromosomes. D. melacanthus has 2n = 12 (10A + XY); L. viridis showed 2n = 14 (12A + XY); and E. collaris showed 2n = 14 (12A + XY). C-banding was performed for the first time in these species and revealed terminal and interstitial heterochromatic regions on the autosomes; DAPI/CMA3 staining showed different fluorescent patterns. In all species, fluorescence in situ hybridization (FISH) with 18S rDNA probe identified signals on one autosomal bivalent, this being the first report of FISH application in the species D. melacanthus and L. viridis. The results obtained add to those already existing in the literature, enabling a better understanding of the meiotic behavior of these insects.

FISH-based karyotyping of Pelmatohydra oligactis (Pallas, 1766), Hydra oxycnida Schulze, 1914, and H. magnipapillata Itô, 1947 (Cnidaria, Hydrozoa)

An account is given of the karyotypes of Hydramagnipapillata Itô, 1947, H.oxycnida Schulze, 1914, and Pelmatohydraoligactis (Pallas, 1766) (Cnidaria, Hydrozoa, Hydridae). A number of different techniques were used: conventional karyotype characterization by standard staining, DAPI-banding and C-banding was complemented by the physical mapping of the ribosomal RNA (18S rDNA probe) and H3 histone genes, and the telomeric (TTAGGG)n sequence by fluorescence in situ hybridization (FISH). We found that the species studied had 2n = 30; constitutive heterochromatin was present in the centromeric regions of the chromosomes; the “vertebrate” telomeric (TTAGGG)n motif was located on both ends of each chromosome and no interstitial sites were detected; 18S rDNA was mapped on the largest chromosome pair in H.magnipapillata and on one of the largest chromosome pairs in H.oxycnida and P.oligactis; in H.magnipapillata, the major rRNA and H3 histone multigene families were located on the largest pair of chromosomes, on their long arms and in the centromeric areas respectively. This is the first chromosomal mapping of H3 in hydras.

Karyotype and chromosomal characteristics of rDNA of Cobitis strumicae Karaman, 1955 (Teleostei, Cobitidae) from Lake Volvi, Greece

The karyotype of Greek cobitid fish Cobitisstrumicae Karaman, 1955, from Lake Volvi, Greece, a representative of one of its two major intraspecific phylogenetic lineages, was analysed by means of sequential Giemsa-staining, C-banding, silver-staining, CMA3 fluorescence banding and also by in situ hybridization (FISH) with rDNA probe. The diploid chromosome number was 2n = 50, karyotype composed of 10 pairs of metacentric to submetacentric and 15 pairs of subtelocentric to acrocentric chromosomes. The nucleolus organizer regions (NORs) as revealed by Ag- and CMA3 staining and FISH were situated in the telomeric region of the fourth submetacentric chromosome pair. The chromosomes contained very low content of C-positive heterochromatin. No heteromorphic sex chromosomes were detected. This first karyotype report for any species of lineage Bicanestrinia Băcescu, 1962 shows a simple karyotype dominated by acrocentric chromosomes and possessing single NOR-bearing chromosome pair. Cytotaxonomic implications of this finding for the taxonomy of the genus Cobitis Linnaeus, 1758 are further discussed.

Karyotypic description of the stingless bee Melipona quinquefasciata Lepeletier, 1836 (Hymenoptera, Meliponini) with emphasis on the presence of B chromosomes

Stingless bees are distributed widely in the tropics, where they are major pollinators of several plant species. In this study, the karyotype ofMeliponaquinquefasciataLepeletier, 1836 was analysed, with emphasis on the presence of B chromosomes. Post-defecating larvae were analysed using Giemsa staining, the C-banding technique, sequential staining with fluorochromes, and FISH. The chromosome number ranged from 2n = 18 to 22 (females) and from n = 9 to 13 (males) due to the presence of 0–4 B chromosomes. This result demonstrates thatM.quinquefasciatahas the same chromosomal number as otherMeliponaIlliger, 1806 species. Considering the A complement, heterochromatin was located only in the pericentromeric region of pair 1. Staining with chromomycin A3(CMA3) and labelling with rDNA probe, indicated that this region corresponded to the nucleolus organising region. The B chromosomes ofM.quinquefasciatacould be found in individuals from different localities, they were completely heterochromatic (C-banding) and uniformly stained by 4’,6-diamidino-2-phenylindole (DAPI). Variations in the number of B chromosomes were detected between cells of the same individual, between individuals of the same colony, and between colonies from different localities.

Karyotype Evolution in Harvestmen of the Suborder Cyphophthalmi (Opiliones)

The morphologically uniform suborder Cyphophthalmi represents a basal group of harvestmen (Opiliones). As such, it plays an important role in the reconstruction of the karyotype evolution within this arachnid order. The cytogenetic analysis of 6 representatives of the suborder Cyphophthalmi, namely Miopsalis sp. (2n = 30; Stylocellidae), Austropurcellia arcticosa (Cantrell, 1980) (2n = 30; Pettalidae), Parapurcellia amatola de Bivort & Giribet, 2010 (2n = 32; Pettalidae), Paramiopsalis aff. ramulosus Juberthie, 1962 (2n = 28; Sironidae), Cyphophthalmus duricorius Joseph, 1868 (2n = 24; Sironidae), and Siro carpaticus Rafalski, 1956 (2n = 52; Sironidae) was performed. Fluorescence in situ hybridization with 18S rDNA probe was used to analyze the distribution of major ribosomal RNA genes in harvestmen. We confront the obtained cytogenetic data with current hypotheses on cyphophthalmid phylogeny to reconstruct their karyotype evolution. We conclude that the ancestral karyotype of harvestmen consisted of 2n = 30 elements with 1 chromosome pair bearing terminal rDNA clusters. The rDNA locus was multiplicated in the evolution of Cyphophthalmi. However, decreases as well as increases in the number of chromosomes have been detected in the karyotype evolution of Cyphophthalmi. Our data thus reveal unexpected diversity in cyphophthalmid karyotypes.