Karotypic characterization of turbot (Scophthalmus maximus) with conventional,fluorochrome and restriction endonuclease-banding techniques

A karyotypic analysis was carried out in turbot (Scophthalmus maximus) using conventional banding techniques, fluorochromes and restriction endonucleasebanding. The standard karyotype of turbot is 2n=44 chromosomes, with 48 chromosome arms. Nucleolar organizer regions, which were localized in the short arm of Chromosome Pair No. 3, showed C-and chromomycin A₃-positive staining. C-bands were mainly located at the centromeres, but were also present in some interstitial and telomeric locations. No differences were revealed among constitutive heterochromatin bands after treatment of fixed chromosomes with fluorochrome staining and restriction endonucleases. Digestion with DdeI restriction enzyme produced a pattern of interstitial banding which suggested some degree of differentiation along the chromosome arms in turbot. Neither variation in chromosome number of arm number nor polymorphism in nucleolar organizer regions was revealed in the turbot analyzed.     

Karyotype analysis of the sea urchinParacentrotus lividus (Echinodermata): evidence for a heteromorphic chromosome sex mechanism

A consistent diploid number of 2n = 36 was determined for the sea urchinParacentrotus lividus from the Gulf of Palermo by analysis of mitotic chromosomes of both early developing embryos and male gonads. The haploid numbern = 18 was determined by counts of spermatocyte bivalents at diakinesis. A heteromorphic chromosome sex mechanism of the XY type is likely present in this species. This is indicated by the occurrence of a chromosomal pair, pair No. 2, which is heteromorphic in both morphology and size in about 50% of the mitotic figures (metaphases and anaphases) of einbryos. In addition, heteromorphism of the same pair of chromosomes occurred during spermatogonial metaphases in the five male specimens investigated here. The detection of a low chromosome number (2n = 36) compared to other echinoids (2n = 42 to 44), a heteromorphic chromosome sex system and the involvement of three chromosome pairs in nucleolar organization (NORs) provide evidence of the specialization of theP. lividus karyotype.     

Cytogenetic studies in green mussel, Perna viridis (Mytiloida: Pteriomorphia), from West Coast of India

Chromosomes of Perna viridis were characterized by karyotype analysis, C-banding and nucleolus organizer regions (NORs). The diploid chromosome number was confirmed as 30 and the karyotype is composed of ten metacentric and five submetacentric chromosome pairs. Constitutive heterochromatin blocks were found on all chromosomal pairs except chromosomal pair 15, which showed uniform staining throughout the entire chromosomes. Silver staining revealed nucleolus organizer regions on telomeric region of four chromosomal pairs, viz. 1, 3, 7 and 11. This is the first comprehensive study undertaken on chromosomes of Perna viridis.     

Karyotype differentiation between two species of carangid fishes,genusTrachurus (Perciformes: Carangidae)

A karyological study ofTrachurus trachurus andT. mediterraneus (Perciformes: Carangidae) was conducted by standard, fluorochrome staining (CMA₃, mithramycin, quinacrine mustard, DAPI), C-, Ag-NOR, and Alu-I banding methods. The karyotypes of both species consisted of 2n = 48 chromosomes, but of different FN:T. trachurus possessed a chromosome complement of 2 metacentric and 46 acrocentric elements, fundamental number (FN) = 50 andT. mediterraneus, a chromosome complement of 4 metacentric, 4 submetacentric, 14 subtelocentric and 26 acrocentric chromosomes, FN = 70. In neither of the two taxa investigated were heteromorphic sex chromosomes observed. The nucleolar organizer region was interstitially located on the long arm of the Ist pair of chromosomes in both species, intermediate inT. mediterraneus and subterminal inT. trachurus. Constitutive heterochromatin was found in nearly all centromeric and telomeric regions inT. trachurus; inT. mediterraneus it formed less intense telomeric and centromeric bands and thin interstitial bands on eight chromosome pairs. In addition, the C-positive material reacted differently to the digestion with endonuclease Alu-l in the two species. The results are discussed and compared with karyological data known for other species of Carangidae.     

Cytogenetics in the sacoglossan Oxynoe olivacea (Mollusca: Opisthobranchia): karyotype, chromosome banding and fluorescent in situ hybridization

Developing embryos and sexually mature follicles of the male portion of ovotestis proved to be a suitable material as a source of cleaving cells for advanced cytological investigations on the sacoglossan species Oxynoe olivacea Rafinesque, 1819 (Mollusca: Opisthobranchia). O. olivacea has a diploid chromosomal number of 30 made up of 15 pairs of which six are metacentric/submetacentric (M/SM), four subtelocentric (ST) and five on the borderline between SM and ST. Correspondingly, 15 bivalents occur in spermatocytes at Metaphase I. Constitutive heterochromatin is scarce and restricted to small C-bands seen in five pachytene bivalents. The use of combined silver staining and fluorescent in situ hybridization (FISH) with 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, revealed that nucleolus organizer regions (NORs) are situated terminally on one arm of a small metacentric pair. The telomeric (TTAGGG) _n sequence did not hybridize with termini of O. olivacea chromosomes. Received: 14 December 1999 / Accepted: 10 July 2000     

Cytogenetic characterization of the greater amberjack, Seriola dumerili (Pisces: Carangidae), by different staining techniques and fluorescence in situ hybridization

A cytogenetic analysis by several staining techniques was carried out on specimens of Seriola dumerili from the Tyrrhenian coast of Sicily. The karyotype was found to be composed of 48 chromosomes in all the specimens examined. All chromosomes are subtelo- and acrocentrics, except for one pair of small-sized submetacentric chromosomes. After Ag-staining, nucleolus organizer regions (NORs) can be observed on the short arms of a medium-sized subtelocentric chromosome pair, and after chromomycin A₃-staining a bright fluorescent signal is evident at the same sites. The 18S rDNA mapping by fluorescence in situ hybridization confirms this unique NOR location. The constitutive heterochromatin is mainly restricted to centromeres and the fluorochrome-staining by chromomycin A₃ and 4′,6-diamidino-2-phenylindole revealed a uniform base composition along the chromosomal arms; therefore stock/population-specific marker chromosomes cannot be identified. Such cytogenetic features, however, rule out the presence of morphologically differentiated sex-chromosomes in the greater amberjack. Received: 16 January 1997 / Accepted: 31 January 1997     

Karyotype analyses reveal inter-individual polymorphism and association of nucleolus-organizer-carrying chromosomes in Capros aper (Pisces: Zeiformes)

Three different karyomorphs with 2n=46, 2n=44 and 2n=42 for Capros aper (L., 1758) (Zeiformes) collected from the Gulf of Lion, near Banyuls-sur-Mer, France, in July 1990 were determined. Karyomorphs were characterized by the same arm number [Fundamental number (FN)=50], suggesting that chromosome variations are due to Robertsonian fusions. In somatic metaphase spreads stained with silver nitrate, nucleolus organizer regions (NORs) consistently occupied a terminal position on the short arms of two small submetacentric chromosomes. Ca. 30% of silver-stained metaphases in each specimen showed NOR chromosomes associated in pairs by their nucleolus organizer regions.     

Cytogenetic analysis of Epinephelus marginatus (Pisces: Serranidae), with the chromosome localization of the 18S and 5S rRNA genes and of the (TTAGGG)n telomeric sequence

 A cytogenetic analysis was carried out on specimens of Epinephelus marginatus (Lowe, 1834) from three localities in the Mediterranean Sea, to deepen knowledge of the chromosome complement of the species and identify any possible population-specific cytogenetic markers. All specimens had a 2n = 48 acrocentric karyotype with two Ag-, chromomycin A₃- and C-positive NORs (nucleolar organizer regions) in the subcentromeric region of the smallest chromosome pair. The constitutive heterochromatin was distributed centromerically. Except for NORs, neither eu- nor heterochromatin show fluorescence after fluorochrome staining, i.e. there is no localized increase of AT- or GC-rich DNA. In all populations, the (TTAGGG) _n telomeric sequences are restricted to the telomeres, and the 18S and the 5S rDNA clusters are located on different chromosome pairs. In specimens from Sardinia, additional signals after C-banding, Ag-staining and FISH (fluorescence in situ hybridization) with 18S rDNA can be observed in the telomeric region of one or two large-sized chromosomes, classified as No. 2. This suggests that additional and variable NORs could be detected in the species in addition to those on the genus-specific, NOR-bearing Chromosome Pair 24. Received: 20 October 1999 / Accepted: 14 April 2000     

Spermatocyte chromosome banding studies inBuccinulum corneum (Prosobranchia: Neogastropoda): Variation in silver-NOR banding pattern

Diploid number (2n=72), and haploid number (n=36) forBuccinulum corneum (L. 1758) collected from the Gulf of Palermo in December 1987 were determined. A simple method to obtain nucleolar organizer regions (NOR), and constitutive heterochromatin regions (C-bands) of chromosomes ofB. corneum is described. Analyses of silver-stained chromosome preparations ofB. corneum suggest that a within-individual variability in NOR-banding pattern is present in each of the five specimens analysed.     

电离辐射对人类染色体的影响

通过对２６例静电加速器工作者染色体的观测，研究电离辐射引起的人类染色体畸变。效应与辐射剂量相关，畸变的类型和频率反映细胞损伤程度。     

Les caryotypes de quelques espèces de bivalves et de gastéropodes marins

The karyotypes of the bivalves Ostrea edulis (2 n=20), Crassostrea gigas (2 n=20), Mytilus galloprovincialis (2 n=28) and M. edulis (2 n=28) were obtained from mitotic metaphases of branchial tissue by means of a cellular suspension technique. The occurrence of metacentric, submetacentric and telocentric chromosomes in O. edulis as well as differences in size between Pair 1 and Pair 10 emphasize the cytological interest of this species compared to C. gigas which possesses only metacentric and submetacentric chromosomes. The variation in Pair 2, which is telocentric in M. galloprovincialis and metacencentric in M. edulis, has been observed for the first time and may support taxological separation of these two species. The karyotypes of the mesogastropods Rissoa ventricosa (2 n=32) and Littorina neritoides (2 n=34) were obtained from mitotic metaphases of gonadal tissue by the same technique. Sexual chromosomes were identified in the karyotypes of R. ventricosa. At the beginning of spermatogenesis, the gonad of L. neritoides shows both haploïd and polyploïd meïotic metaphases which correspond to typical (germinal cells) and atypical (nurse cells) forms, respectively.     

Genetic population structure of a species' complex of blue mussels (Mytilus spp.)

Blue mussels representing two nominal species (Mytilus trossulus Gould, 1850 and Mytilus galloprovincialis Lamarck, 1819 were collected from 28 intertidal locations along the Pacific coast of the USA in 1990–1991 (total N=1255) and examined for variation at 15 allozyme loci. Twelve samples, mostly from a region of suspected hybridization, were analyzed for variation in seven shell characters. Principal-components analysis of allozyme data revealed three groups based on first principal-component scores, which were identified as M. trossulus, M. galloprovincialis, and hybrids. Canonical discriminant analysis of shell characters was less successful in separating mussels into discrete groups. Each location was characterized for four environmental variables: (1) temperature, (2) salinity, (3) tidal height and (4) degree of exposure to wave action, which were then used as independent variables in a series of multiple-regression analyses, with the proportions of the two species as dependent variables. Temperature and salinity had significant (P<0.05) effects on the macrogeographic distribution of the two species, whereas the effects of height in the tidal zone and degree of wave exposure were not statistically significant. Salinity was found to have a greater influence than temperature on the microgeographic distribution of the two species. M. trossulus was more abundant at locations with lower temperatures and greater salinity variation than M. galloprovincialis. The two species appear to be ecologically distinct, and their genetic integrity is at least partly the result of environmental heterogeneity.     

Number and morphology of chromosomes in three geographical populations of Ascidiella aspersa (Ascidiacea)

In contrast with some previous reports, we have found the same number of chromosomes (n=9, 2n=18) for Ascidiella aspersa Müller from the Gulf of Naples, the west coast of Scotland and the Lagoon of Venice. It follows that the hypothesis, that different geographical populations of A. aspersa might possess different chromosome numbers, is no longer tenable. The karyogram of A. aspersa shows 9 pairs of metacentric autosomes. The oocyte bivalents show chiasmata and are characterized by the faintness with which they are stained.This investigation was supported by a grant from C.N.R. for Marine Biology, n. 69.00117 14.43.3.     

Karyology of the toadfish Porichthys plectrodon (Jordan and Gilbert, 1882) (Batrachoididae) from Margarita Island,Venezuela

This paper reports the results of cytogenetic analyses carried out on Porichthys plectrodon using conventional Giemsa staining, C-banding and silver staining techniques. A diploid chromosome count of 2n=44 was observed, consisting of 8 metacentric, 10 submetacentric, 6 subtelocentric and 20 acrocentric chromosomes. Differences in length made it possible to identify homologous chromosomes within the metacentric group. Constitutive heterochromatin was distributed as large pericentromeric blocks in pairs 1 and 2, while the rest of the chromosomes were marked in centromeric regions, some more conspicuously than others. One pair of small-sized acrocentric NOR-bearing chromosomes (21) was identified by the nucleolar regions located terminally on their short arms.Communicated by P.W. Sammarco, Chauvin     

Karyotype and sex determination in <Emphasis Type="Italic">Dinophilus gyrociliatus</Emphasis> (Polychaeta: Dinophilidae)

The study of the male and female karyotypes of the polychaete Dinophilus gyrociliatus was performed with the aim of ascertaining the diploid number of chromosomes and confirming the existence of karyological differences between the two sexes. Our results evidenced that the female karyotype of D. gyrociliatus consists of 2n=24 chromosomes, while the male karyotype has 2n=23 chromosomes. X chromosomes are subtelocentric and much bigger than the autosomes. The size of the D. gyrociliatus chromosomes, which in the female embryos vary in length from little more than 2.8 wm to less than 0.7 wm, are relatively small by comparison with those of other polychaetes. The existence of maternal and environmental factors able to influence the sex ratio of D. gyrociliatus and chromosomal differences between the sexes confirms that: (1) sex determination is chromosomal and syngamic of the XX-X0 type and (2) control of the sex ratio is progamic and depends on genetic and environmental factors.     

Karyotypic characterization of the great sturgeon, Huso huso, by multiple staining techniques and fluorescent in situ hybridization

A karyotype analysis by several staining techniques was carried out on the great sturgeon, Huso huso (Linnaeus, 1758). The karyotype (2n = 118 ± 2) was composed of 42 pairs of meta-/submetacentric chromosomes and 17 pairs of acrocentrics/microchromosomes. Constitutive heterochromatin was mainly located at the centromeric regions of the acrocentric chromosomes. The biarmed chromosomes showed weak C-bands. Fluorescent staining with GC-specific chromomycin A₃ showed clearly recognizable fluorescent regions, whereas a more uniform base composition was revealed by the AT-specific 4,6-diamidino-2 phenylindole. After Ag-staining, nucleolar organizer regions could be observed on the short arms of two medium-sized submetacentrics and on two acrocentrics. Digoxigenated 28S and 5S rDNA probes, prepared from Acipenser naccarii DNA and hybridized to metaphase chromosomes, showed signals on six and two chromosomes, respectively. The telomeric sequence (TTAGGG) _n detected by FISH was located at both ends of each chromosome. Results are discussed in relation to karyotype organization and evolution in sturgeons. Received: 1 April 1998 / Accepted: 21 July 1998     

Cytogenetic analysis of Oedalechilus labeo (Pisces: Mugilidae), with a report of NOR variability

 A cytogenetic analysis by several staining techniques was carried out on Oedalechilus labeo specimens from the west coast of Italy, in order to extend the karyological knowledge on Mediterranean mullets. The karyotype of the species is composed of 46 acrocentric and 2 subtelocentric chromosomes. Constitutive heterochromatin was found to be restricted to the centromeres. Fluorochrome staining revealed a uniform base composition along the chromosome arms. Nucleolus organizer regions (NORs) are located on the short arms of chromosome pair 9 and can be either homomorphic or quite heteromorphic in size. In some specimens chromomycin A₃-staining and fluorescence in situ hybridization with 18S rDNA revealed a third additional and inactive NOR on the short arms of a medium-sized chromosome. This is the first report of NOR variability in Mugilidae, a family generally characterized by a quite conservative karyotype. Heterochromatin composition and NOR location differentiate O. labeo from a cytotaxonomical point of view from the Liza species studied, Liza being considered the genus from which the genus Oedalechilus derived. Received: 10 April 1999 / Accepted: 21 October 1999     

Replication,C and Ag-NOR chromosome-banding in two anguilliform fish species

The chromosomes of the Conger conger and Echelus myrus (Anguilliformes: Teleostei), were analysed by conventional (C and Ag-NOR) and by replication-banding techniques. Both species had a diploid number of 38 and one chromosome pair with C-positive nucleolar organizer regions (NOR). These cytogenetic traits are mainly consistent with those recorded for anguilliform species in general. The replication-banding pattern allowed us to identify individual chromosome pairs, to detect different classes of chromatin, and to suggest chromosome evolutive mechanisms in these two species. The results confirm the superiority of replication patterns in resolving longitudinal differentiation of fish chromosomes, that is not generally possible from structural bands.     

Seasonal variation in movement,aggregation and destructive grazing of the green sea urchin (<Emphasis Type="Italic">Strongylocentrotus droebachiensis</Emphasis>) in relation to wave action and sea temperature

Hydrodynamic forces are an important determinant of subtidal community structure, particularly when they limit the distribution and foraging ability of mobile consumers. We examined the effect of wave action on the rate of movement and destructive grazing of a kelp bed by the green sea urchin (Strongylocentrotus droebachiensis) under field conditions. We measured density and rate of advance at fixed intervals along ∼100 m of a grazing front over 1 year, and quantified individual movement rates in the barrens 5–10 m behind the urchin front using a time-lapse videography. Seasonal variation in the mean rate of advance of the front (range: 0–4 m month⁻¹) was explained by changes in urchin density at the front (120–360 individuals m⁻²), which in turn varied inversely with significant wave height (0.5–2 m). Water temperature (0.8–17.6°C) had no effect on the rate of advance or on urchin density (aggregation) at the front, except when temperature exceeded 17°C. Movement of individual urchins also was affected by wave action: we observed a significant decrease in speed and displacement of urchins with increasing significant wave height. Wave action had no effect on the proportion of urchins moving or the degree of linearity of their movements. We propose that the decrease in urchin density at the front associated with increased wave action, results from de-aggregation, which reduces the risk of dislodgement, combined with a reduction in urchin movement in barrens, which supplies new urchins to the front.     

Abundance,population structure and claw morphology of the semi-terrestrial crab <Emphasis Type="Italic">Pachygrapsus marmoratus</Emphasis> (Fabricius, 1787) on shores of differing wave exposure

Wave action is known to influence the abundance and distribution of intertidal organisms. Wave action will also determine the duration and suitability of various foraging windows (high-tide and low-tide, day and night) for predation and can also affect predator behaviour, both directly by impeding prey handling and indirectly by influencing prey abundance. It remains uncertain whether semi-terrestrial mobile predators such as crabs which can access intertidal prey during emersion when the effects of wave action are minimal, are influenced by exposure. Here, we assessed the effect of wave action on the abundance and population structure (size and gender) of the semi-terrestrial intertidal crab Pachygrapsus marmoratus on rocky shores in Portugal. The activity of P. marmoratus with the tidal cycle on sheltered and exposed shores was established using baited pots at high-tide to examine whether there was activity during intertidal immersion and by low-tide searches. Because prey abundance varies along a wave exposure gradient on most Portuguese shores and because morphology of crab chelipeds are known to be related to diet composition, we further tested the hypothesis that predator stomach contents reflected differences in prey abundance along the horizontal gradient in wave exposure and that this would be correlated with the crab cheliped morphology. Thus, we examined phenotypic variation in P. marmoratus chelipeds across shores of differing exposure to wave action. P. marmoratus was only active during low-tide. Patterns of abundance and population structure of crabs did not vary with exposure to wave action. Stomach contents, however, varied significantly between shores of differing exposure with a higher consumption of hard-shelled prey (mussels) on exposed locations, where this type of prey is more abundant, and a higher consumption of barnacles on sheltered shores. Multivariate geometric analysis of crab claws showed that claws were significantly larger on exposed shores. There was a significant correlation between animals with larger claws and the abundance of mussels in their stomach. Variation in cheliped size may have resulted from differing food availability on sheltered and exposed shores.