Maria Stella Colomba

Ribosomal DNA Location in the Scarab Beetle Thorectes Intermedius (Costa) (Coleoptera: Geotrupidae) Using Banding and Fluorescent in-situ Hybridization

Mitotic metaphase chromosomes of the scarab beetle Thorectes intermedius (Costa) (Coleoptera Scarabaeoidea: Geotrupidae) were analyzed using various banding methods and fluorescent in-situ hybridization (FISH) with a ribosomal probe. The results obtained indicate that silver and CMA3 staining are unable to localize the chromosome sites of nucleolar organizer regions (NORs). Such an inadequacy is a consequence of the extensive silver and CMA3 stainability of both constitutive heterochromatin and heterochromatin associated to the NORs.

FISH mapping of 18S-28S and 5S ribosomal DNA, (GATA) n and (TTAGGG) n telomeric repeats in the periwinkle Melarhaphe neritoides (Prosobranchia, Gastropoda, Caenogastropoda)

Spermatocyte chromosomes of Melarhaphe neritoides (Mollusca, Prosobranchia, Caenogastropoda) were studied using fluorescent in situhybridization (FISH) with four repetitive DNA probes (18S rDNA, 5S rDNA, (TTAGGG)n and (GATA)n). Single-colour FISH consistently mapped one chromosome pair per spread using either 18S or 5S rDNA as probes. The telomeric sequence (TTAGGG)n hybridized with termini of all chromosomes whereas the (GATA)n probe did not label any areas. Simultaneous 18S-5S rDNA and 18S-(TTAGGG)n FISH demonstrated that repeated units of the three multicopy families are closely associated on the same chromosome pair.

Cytogenetic characterization of Brachidontes pharaonis (Fisher P., 1870): Karyotype, banding and fluorescent in situ hybridization (fish) (Mollusca: Bivalvia: Mytilidae)

Abstract The mussel Brachidontes pharaonis (Fisher P., 1870) (Bivalvia: Mytilidae) has a diploid chromosomal set of 28 made up of 14 pairs of which eight are mono-armed (ST) and six bi-armed (M+SM). Fourteen bivalents occur in spermatocytes both at pachytene and metaphase-I. The use of combined silver and CMA3 staining reveals that nucleolus organizer regions (NORs) are located terminally on the long arm of a small subtelocentric chromosome pair (pair 14) and are compartmentalized in GC base pairs. A Paracentrotus lividus (Echinodermata) 4.3 kilobase (kb) rDNA probe (prR14) consisting of sequences from the 3′ end of 18S rDNA to the 3′ end of 26S rDNA was used to map the rDNA loci of B. pharaonis by fluorescence in situ hybridization. The results obtained with this technique confirm that NORs are located terminally on two small subtelocentric chromosomes (pair 14) and establish that the difference in dimension of homologous NORs is due to difference in the number of rDNA copies.

Chromosomal Location of Ribosomal DNA (rDNA), (GATA)n and (TTAGGG)n Telomeric Repeats in the Neogastropod Fasciolaria Lignaria (Mollusca: Prosobranchia)

This paper reports on a successful application of fluorescent in situhybridization (FISH) with three repetitive DNA probes (ribosomal DNA (rDNA), (GATA)nand (TTAGGG)n) in the chromosomes of Fasciolaria lignaria(Mollusca: Prosobranchia: Neogastropoda). rDNA FISH consistently identified four chromosome pairs per spread in the three examined specimens. The telomeric sequence (TTAGGG)nhybridized with termini of all chromosomes. GATA FISH revealed abundant, dispersed minisatellite regions which were not associated to the XY sex-determining mechanism as indicated by the absence of a Y specific pattern of labelling.

Physical mapping of rDNA genes, (TTAGGG)n telomeric sequence and other karyological features in two earthworms of the family Lumbricidae Annelida: Oligochaeta)

A cytogenetical study was carried out on the chromosomes and nuclear DNA amounts of the terrestrial earthworms Octodrilus complanatus and Eisenia foetida (Annelida: Oligochaeta: Lumbricidae). Chromosomes were studied using Giemsa staining, banding methods and fluorescent in situ hybridization (FISH) with two repetitive DNA probes [rDNA and (TTAGGG)n]. rDNA FISH and silver staining consistently identified one chromosome pair per spread in both species. The telomeric sequence (TTAGGG)n hybridized with termini of all the chromosomes in both earthworms. Flow cytometry DNA assays showed that O. complanatus and E. foetida had different nuclear DNA contents (2C value=1.72 and=1.40 pg, respectively) but very similar base composition in their genomes.

High heterochromatin content in somatic chromosomes of two unrelated species of Diplopoda (Myriapoda)

For the first time, a conventional analysis of C-banded karyotypes was carried out in two distantly related diplopod species; this revealed an impressive percentage of heterochromatin in both genomes. In Acanthopetalum sicanum (Order Callipodida) (2n = 12), heterochromatin constitutes about 60% of the total DNA in females and 56% in males, whereas in Enologus oxypygum (Order Julida) (2n = 22) it is about 67% in both sexes. Heterochromatin of the two species was found to be similar in base composition (AT rich) and heterochromatin distribution, indicating that it has accumulated in a species-specific manner. Sex-determining mechanisms of the XY type were detected in both A. sicanum and E. oxypygum. In A. sicanum, the Y presented the lowest heterochromatic content of all chromosomes in the karyotype, whereas the X presented the highest.

Multiple-chromosome sex systems in the darkling beetles Blaps gigas and Blaps gibba (Coleoptera, Tenebrionidae)

We have studied mitotic and meiotic chromosomes in the males of two species of Blaps: B. gigas and B. gibba. Karyological characteristics such as the occurrence of a multivalent configuration at diakinesis and two types of metaphase-II spreads support the notion that multiple-chromosome sex systems involving five chromosomes in B. gigas and eight chromosomes in B. gibba have developed in these species. Results obtained by means of silver staining and C-banding techniques suggest that the complex sex systems occurring in B. gigas and B. gibba may have originated from exchanges of terminal ribosomal genes among the Y chromosome and some autosomes.

Characterization of mitotic and meiotic, chromosomes of the vermetid gastropod Dendropoma (Novastoa) Petraeum (Monterosato, 1884) (Mollusca, Caenogastropoda)

Abstract The vermetid gastropod Dendropoma (Novastoa) petraeum from the Northwestern coast of Sicily was investigated karyologically. The count of spermatocyte bivalents at diakinesis gave the haploid number of this species as n=17. The count of mitotic chromosomes in males, females and early developing embryos gave the diploid number as 2n=34. Irrespective of sex, the karyotype of D. petraeum consists of 17 homomorphic chromosome pairs, of which 8 are metacentric, 4 subtelocentric and 5 acrocentric. This species is characterized by the presence of two chromosomes per cell involved in nucleolus organization (NOR) and by a low amount of constitutive heterochromatin.

Chromosomes of the Parasitic Isopod Anilocra Physodes

Counts of mitotic chromosomes have allowed to determine 2n = 12 as the modal diploid number of Anilocra physodes. Application of the C-banding technique reveals that two hetero-chromatic blocks bordering the centromeric region occur in each chromosome. This supports the notion that the actual low chromosome number of A. physodes may have been derived through a process of Robertsonian fusion.