Anna Rita Rossi

Cytogenetic analysis of global populations of Mugil cephalus (striped mullet) by different staining techniques and fluorescent in situ hybridization

The present paper reports the results of cytogenetic analysis carried out on several scattered populations of the striped mullet, Mugil cephalus, the most widespead among mugilid species. The karyotype was investigated through Ag-staining, C-banding, fluorochrome-staining (chromomycin A3/DAPI) and fluorescent in situ hybridization with rDNA genes. All populations showed the same chromosome number and morphology and no changes were detected in heterochromatin and NORs. Therefore, neither population- nor sex-specific marker chromosomes were identified. In some of the specimens, NOR size heteromorphism was detected. Results are discussed with respect to karyotype and ribosomal cistrons organization and to cytotaxonomic implications.

Cytogenetic analysis of Liza ramada (Pisces, Perciformes) by different staining techniques and fluorescent in situ hybridization

A cytogenetic investigation was carried out on specimens of Liza ramada, a mugilid species common in the Mediterranean sea. The analysis of chromosomes was performed through Ag-staining, C-banding, chromomycin A3 and DAPI staining, and fluorescent in situ hybridization with ribosomal genes. The results obtained are discussed with respect to cytotaxonomic implications and to the features of NORs.

Phylogenetic Analysis of Mediterranean Mugilids by Allozymes and 16S mt-rRNA Genes Investigation: Are the Mediterranean Species of Liza Monophyletic?

The family Mugilidae (Pisces, Mugiliformes) includes species which are present in all tropical and temperate regions. Six species, Chelon labrosus, Mugil cephalus, Liza aurata, L. ramada, L. saliens, Oedalechilus labeo, are commonly found in the Mediterranean. These species have been widely studied through morphological, biochemical, and molecular markers. However, their phylogenetic relationships, and therefore the assumed monophyly of Liza species, still remain unclear. To further investigate this topic, gene–enzyme systems and sequences of the partial 16S rRNA mitochondrial gene were analyzed in Italian samples of all six Mediterranean species. The phylogenetic reconstructions indicated M. cephalus as being the most divergent species and the existence of a main cluster including all the Mediterranean species of Liza and C. labrosus. The parametric bootstrap approach adopted to test alternative phylogenetic hypotheses indicated that the Mediterranean species of Liza do not form a monophyletic group exclusive of Chelon.

Geographic variability in the grey mullet Mugil cephalus: preliminary results of mtDNA and chromosome analyses

Abstract The grey mullet, Mugil cephalus , plays an important role in the fisheries and aquaculture of tropical and subtropical regions of the world. This species is considered cosmopolitan, but its distribution appears peculiar with regard to the coastal ecology of the species. A multidisciplinary study of the geographic variability of this species, through a cytogenetic, molecular and morphometric characterization, was undertaken to detect whether genetically distinct populations occur. The preliminary results from analyses of mitochondrial DNA and of chromosomes of seven different populations around the world are reported. The different populations analysed are well discriminated by mtDNA analyses: samples are clustered in four groups, Mediterranean, East Atlantic, Central Pacific and East Pacific, with a maximum sequence divergence of 3.3%. The karyotype of all the populations studied is uniformly composed of 48 acrocentric chromosomes, and neither heterochromatin distribution nor nucleolus organizer regions allow the identification of chromosomal markers useful in distinguishing these genetically differentiated groups of populations.

Comparative molecular cytogenetic analysis of two congeneric species, Mugil curema and M. liza (Pisces, Mugiliformes), characterized by significant karyotype diversity

Two congeneric mullet species, Mugil liza and M. curema, respectively with an all-uniarmed and an all-biarmed karyotype, were cytogenetically studied by base-specific fluorochrome staining and FISH-mapping of 45S and 5S ribosomal RNA genes (rDNA) and the (TTAGGG)n telomeric repeats. Whereas 45S rDNA sites might be homeologus in the two species, 5S rDNA sites are not, as they are localized on chromosome arms of different size. In both species, the (TTAGGG)n telomeric probe hybridized to natural telomeres and was found scattered along the NORs. In metacentric chromosomes of M. curema, no pericentromeric signals of the telomeric probe were detected. Data are discussed in relation to the karyotype evolution in Mugilidae and to the mechanisms and the evolutionary implications of Robertsonian rearrangements in M. curema.

Genetic structure of gilthead seabream, Sparus aurata, in the Central Mediterranean Sea

The gilthead seabream, Sparus aurata, represents an important economic resource for Mediterranean aquaculture. In spite of its wide geographic distribution and economic importance, only recently studies have been carried out on the genetic composition of natural populations, which have revealed a picture of a heterogeneous degree of genetic differentiation among S. aurata populations. In this study an allozyme analysis of samples from six different collecting sites along the Italian and Croatian coasts was carried out, covering an area in the Central Mediterranean sea that has yet to be investigated through gene-enzyme systems. Data on 26 gene loci, 10 of which are polymorphic, indicate a slight but significant genetic structure (FST = 0.0167) of the species. A hierarchical analysis of population subdivision made it possible to identify three different assemblages found in the Adriatic Sea, Tyrrhenian Sea and Sardinian Channel, though an isolation by distance model can be rejected. The results are discussed in the light of previous literature and taking conservation into consideration.

Testing Species Delimitations in Four Italian Sympatric Leuciscine Fishes in the Tiber River: A Combined Morphological and Molecular Approach

Leuciscine fishes represent an important component of freshwater ichthyofauna endemic to northern Mediterranean areas. This lineage shows high intra-specific morphological variability and exhibits high levels of hybridization, two characteristics that contribute to systematic uncertainties, misclassification of taxa and, potentially, the mismanagement of biodiversity. This study focused on brook chub, Squalius lucumonis, an endemic taxon of Central Italy. The taxonomic status of this species has long been questioned, and a hybrid origin from sympatric leusciscines (S. squalus x Rutilus rubilio, or S. squalus x Telestes muticellus) has been hypothesised. A phenotypic (evaluating shape and meristic counts) and genetic (using mitochondrial and nuclear markers) investigation of these four taxa was conducted to test species delimitation in sympatric areas and to evaluate the taxonomic status of S. lucumonis. One hundred and forty-five individuals of all four taxa were collected within streams of the lowest portion of the Tiber River basin and analysed; this region encompasses a large portion of the S. lucumonis distribution. The different morphological and genetic approaches were individually examined, compared, and then combined in a quantitative model to both investigate the limits of each approach and to identify cases of misclassification. The results obtained confirm the cladogenetic non-hybrid origin of S. lucumonis, highlight the need for immediate conservation actions and emphasise the value of an integrated approach in the study of leuciscines evolution.

Karyotype characterization of Mugil incilis Hancock, 1830 (Mugiliformes: Mugilidae), including a description of an unusual co-localization of major and minor ribosomal genes in the family

This study reports the description of the karyotype of Mugil incilis from Venezuela. The chromosome complement is composed of 48 acrocentric chromosomes, which uniformly decrease in size. Therefore, the homologues can not be clearly identified, with the exception of one of the largest chromosome pairs, classified as number 1, whose homologues may show a subcentromeric secondary constriction, and of chromosome pair number 24, which is considerably smaller than the others. C-banding showed heterochromatic blocks at the centromeric/pericentromeric regions of all chromosomes, which were more conspicuous on chromosomes 1, given the C-positive signals include the secondary constrictions. AgNO3 and fluorescent in situ hybridization (FISH) with 45S rDNA demonstrated that the nucleolus organizer regions are indeed located on the secondary constrictions of chromosome pair number 1. FISH with 5S rDNA revealed that the minor ribosomal genes are located on this same chromosome pair, near the NORs, though signals are closer to the centromeres and of smaller size, compared to those of the major ribosomal gene clusters. This is the first description of co-localization of major and minor ribosomal genes in the family. Data are discussed from a cytotaxonomic and phylogenetic perspective.

Cytogenetic characterization of Rhomboplites aurorubens and Ocyurus chrysurus, two monotypic genera of Lutjaninae from Cubagua Island, Venezuela, with a review of the cytogenetics of Lutjanidae (Teleostei: Perciformes)

Lutjanidae, commonly known as snappers, includes 105 species, grouped in four subfamilies. In spite of the high number of species and of its worldwide distribution, the family has been little investigated and the phylogenetic relationships among some of its genera and species are still cause for debate. Only a small number of the species has been cytogenetically analysed. This study reports the first description of the karyotype of Rhomboplites aurorubens as well as data concerning the distribution of the constitutive heterochromatin and the location of the 18S rRNA and the 5S rRNA genes. Specimens of Ocyurus chrysurus from Venezuela were also investigated for the same cytogenetic features. Both species have a 48 uniarmed karyotype, but R. aurorubens has a single subtelocentric chromosome pair, the smallest of the chromosome complement, among the other acrocentric chromosomes. The C-positive heterochromatin is limited to the pericentromeric regions of all chromosomes. Both species show a single chromosome pair bearing the Nucleolus Organizer Regions, but NORs are differently located, in a terminal position on the short arms of the smallest chromosomes in R. aurorubens and in a paracentromeric position in a chromosome pair of large size in O. chrysurus. In O. chrysurus, the 5S rDNA gene cluster is located on a medium-sized chromosome pair, whereas in R. aurorubens it is syntenic with the 18S rDNA gene cluster on chromosome pair number 24. The obtained cytogenetic data, along with previous cytogenetic, morphological and molecular data for the family, reinforce the proposal to synonymize genus Ocyurus with Lutjanus. A review of Lutjanidae cytogenetics is also included.

Cytogenetic studies in three species of Lutjanus (Perciformes: Lutjanidae: Lutjaninae) from the Isla Margarita, Venezuela

In the present study, three species of Lutjaninae, Lutjanus analis, L. griseus and L. synagris, were analyzed by conventional Giemsa staining, C-banding and silver staining, to reveal active Nucleolus Organizer Regions (NORs). Fluorescent in situ hybridization (FISH) was also applied to establish the number and location of the ribosomal gene clusters (18S and 5S rRNA genes). Counts of diploid metaphasic cells revealed a diploid modal chromosome complement composed of 48 acrocentric chromosomes in both L. analis and L. griseus. Two cytotypes were observed in L. synagris: cytotype I, with 2n=48 acrocentric chromosomes, found in 19 specimens, and cytotype II, with 46 acrocentric chromosomes and one large metacentric, found in two specimens. The large metacentric, which possibly originated from a Robertsonian rearrangement, was not found to be sexrelated. In the three species, constitutive heterochromatin is located in the centromeres of all chromosomes. NORs were detected on the short arms of a single chromosome pair, number 24 in L. analis and number 6 in both cytotypes of L. synagris. In L. griseus, a polymorphism of the NORs number was detected, by both Ag-staining and FISH, as females show a maximum of three NORs, and males a maximum of six NORs. In all species, minor ribosomal genes were found located on a single chromosome pair. The obtained data, along with those previously reported for other five Lutjanidae species, show that a general chromosome homogeneity occurs within the family, but that derived karyotypes based on Robertsonian rearrangements as well as multiple and variable NORs sites can also be found. No presente estudo tres especies de Lutjaninae, Lutjanus analis, L. griseus e L. synagris foram analisadas atraves da coloracao convencional com Giemsa, banda C e coloracao com nitrato de prata para identificar as Regioes Organizadoras de Nucleolo (NORs) ativas. Hibridacao fluorescente in situ (FISH) foi tambem aplicada para estabelecimento do numero e localizacao dos agrupamentos de genes ribossomicos (18S e 5S rRNA). A contagem de celulas metafasicas revelou um numero diploide modal de 48 cromossomos acrocentricos em L. analis e L. griseus. Dois citotipos foram observados em L. synagris: citotipo I com 2n=48 cromossomos acrocentricos, encontrado em 19 especimes, e citotipo II com 46 cromossomos acrocentricos e um grande metacentrico, encontrado em dois especimes. O grande metacentrico, que possivelmente se originou por um rearranjo Robertsoniano, nao esta relacionado com o sexo. Nas tres especies a heterocromatina constitutiva esta localizada nas regioes centromericas de todos os cromossomos. NORs foram detectadas no braco curto de um unico par cromossomico, numero 24 em L. analis e numero 6 em ambos os citotipos de L. synagris. Em L. griseus, um polimorfismo de numero de NORs foi observado, apos coloracao com prata e por FISH, as femeas apresentaram um maximo de tres NORs e os machos um maximo de seis NORs. Em todas as especies os genes ribossomicos 5S foram encontrados em um unico par cromossomico. Os dados obtidos, somados aos demais previamente publicados para cinco outras especies de Lutjanidae, mostram que na familia ha uma homogeneidade cromossomica, porem tambem sao encontrados cariotipos derivados, originados por rearranjos Robertsonianos, assim como pela ocorrencia de sitios multiplos e variados de NORs.