Stick insects have been studied mainly for non-conventional reproduction modes, such as parthenogenesis, hybridogenesis and androgenesis. Parallel karyological investigations have evidenced extensive numerical and structural chromosome re-patterning, particularly evident in hybrid parthenogenetic taxa. Chromosome sets of bisexual Leptynia (Pantel) species show an evolutionary trend from 40 to 36 chromosomes and are characterized by cytological satellites of variable size and localization. We performed fluorescence in situ hybridization (FISH) analysis using 45S ribosomal genes and pentameric (TTAGG)n telomere sequences as probes in two strictly related but karyotypically distinct species, L. montana Scali (2n = 38/37; XX/XO) and L. attenuata Pantel (2n = 36). L. attenuata has recently been split into three subspecies (L. attenuata attenuata, L. attenuata iberica and L. attenuata algarvica), and found to share an XX/XY sex chromosome formula, unusual for stick insects. FISH by 45S rDNA sequences consistently labelled the short arm of the 4th chromosome pair, often of a variable size. Silver staining showed that nucleolar organizer regions (NORs) are active. FISH of the telomeric repeats, besides ordinary telomeres, also labelled the short arm of this same pair. The use of both probes in double FISH analysis fully confirmed the co-localization of ribosomal and telomeric highly repeated sequences. Since it is increasingly emerging that the co-localization of NORs and telomeric sequences appears to be a feature shared by evolutionarily distant animals, its possible role is discussed.

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The Muraenidae is one of the largest and most complex anguilliform families. Despite their abundance and important ecological roles, morays are little studied, especially cytogenetically, and both their phylogenetic relationships and the taxonomy of their genera are controversial. With the aim of extending the karyology of this fish group, the chromosomal mapping of the 5S ribosomal gene family was performed on seven species belonging to the genera Muraena and Gymnothorax from both the Atlantic and Pacific oceans. Fluorescence in situ hybridisation (FISH) experiments were realized using species-specific 5S rDNA probes; in addition, two-colour FISH was performed to investigate the possible association with the 45S ribosomal gene family. Multiple 5S rDNA clusters, located either in species-specific or in possibly homoeologous chromosomes, were found. Either a syntenic or different chromosomal location of the two ribosomal genes was detected. Our results revealed variability in the number and location of 5S rDNA clusters and confirmed a substantial conservation of the number and location of the 45S rDNA.

A comparative cytogenetic analysis, using both conventional staining techniques and fluorescence in situ hybridization, of six Indo-Pacific moray eels from three different genera (Gymnothorax fimbriatus, Gymnothorax flavimarginatus, Gymnothorax javanicus, Gymnothorax undulatus, Echidna nebulosa and Gymnomuraena zebra), was carried out to investigate the chromosomal differentiation in the family Muraenidae. Four species displayed a diploid chromosome number 2n = 42, which is common among the Muraenidae. Two other species, G. javanicus and G. flavimarginatus, were characterized by different chromosome numbers (2n = 40 and 2n = 36). For most species, a large amount of constitutive heterochromatin was detected in the chromosomes, with species-specific C-banding patterns that enabled pairing of the homologous chromosomes. In all species, the major ribosomal genes were localized in the guanine-cytosine-rich region of one chromosome pair, but in different chromosomal locations. The (TTAGGG)n telomeric sequences were mapped onto chromosomal ends in all muraenid species studied. The comparison of the results derived from this study with those available in the literature confirms a substantial conservation of the diploid chromosome number in the Muraenidae and supports the hypothesis that rearrangements have occurred that have diversified their karyotypes. Furthermore, the finding of two species with different diploid chromosome numbers suggests that additional chromosomal rearrangements, such as Robertsonian fusions, have occurred in the karyotype evolution of the Muraenidae.