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.     

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 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.     

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     

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 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     

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     

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.     

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.     

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 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     

Karyotype, nucleolus organiser regions and constitutive heterochromatin in Ostrea angasi (Molluscae: Bivalvia): evidence of taxonomic relationships within the Ostreidae

Chromosome preparations from gill tissue of the Australian flat oyster Ostrea angasi Sowerby were studied with conventional Giemsa staining, C-banding and silver staining techniques. A diploid complement of 2n = 20 was observed, consisting of five metacentric, three submetacentric and two subtelocentric pairs. Constitutive heterochromatin was distributed as large centromeric blocks in Chromosome Pairs 3, 6, 8, 9 and 10. The nucleolus organizer regions (NORs) were located terminally on long arms of Chromosome Pairs 9 and 10. This allowed the identification of homologous chromosomes in submetacentric and subtelocentric pairs. Intraspecific variability in NOR pattern as revealed by differences in the number of silver-stained NORs (Ag-NORs) per cell was found to be very common. Comparison of the patterns of karyotype, C-band and Ag-NORs between species of the larviparous oysters for which data have been published demonstrate that the chromosomal structure of the endemic Australian and New Zealand species O. angasi shows little similarity to the Southern Hemisphere oysters O. (Eostrea)puelchana Orbigny and Tiostrea chilensis (Philippi) and the Indo-West Pacific oyster O. denselamellosa, but very high resemblance to the European species O. edulis. Received: 12 August 1996 / Accepted: 16 September 1996     

Banding pattern of mussel (Mytilus galloprovincialis) chromosomes induced by 2 × SSC/Giemsa-stain treatment

Mytilus galloprovincialis were collected from an intertidal population in NW Spain in 1988, and chromosomes from the gill tissue of 37 individuals were studied. The present paper describes the banding pattern of aM. galloprovincialis population which enabled us to identify all pairs of chromosomes. Banding was induced by means of a 2 × SSC (0.3M sodium chloride:0.03M sodium citrate)/Giemsa-staining technique, and a diagrammatic representation was constructed based on mean number of bands. The application of this banding technique may prove useful in future research related to the cytogenetics and cytotaxonomy of mussels and other molluscs.     

Multiple sex-chromosome system and other karyological characterizations of Pterotrachea hippocampus (Mollusca: Mesogastropoda)

Two modal diploid numbers of chromosomes were found for Pterotraches hippocampus Philippi (Mollusca: Mesogastropoda) collected from the Gulf of Palermo in 1990: 2n=31 and 32 for males and females, respectively. This, along with other karyological characteristics such as the occurrence of a trivalent configuration at diakinesis and two types of metaphase-II spreads in spermatocytes, supports the notion that a X₁X₂Y/X₁X₁X₂X₂ sex mechanism operates in the species investigated here. Silver nitrate procedure revealed an intraindividual variation in the Ag-staining pattern occurring in this species. The majority of the chromosome pair displayed terminal and/or interstitial heterochromatic blocks after C-banding.     

Evaluation of the mutagenic potential of chlorpyrifos (CPF) using polytene chromosomes of Anopheles mosquito

Diverse cytogenetic tests are employed for short term screening of suspect environmental mutagens by using insects and mammals as models. In the present paper the polytene chromosomes of a mosquito Anopheles maculatus were used to evaluate the mutagenic potential of a widely used organophosphate pesticide chlorpyrifos-[o, o-diethyl-o-(3, 5, 6-trichloro-2-pyridyl) phosphothioate]. The results are based on the frequency of various structural aberrations encountered in the polytene chromosomes of the larvae treated with LC20 of chlorpyrifos (CPF). These aberrations were dominated by inversions, stickiness of the chromosomes, heterochromatinization of the bands and lack of polyteny. The frequency of various aberrations was highest in the left arm of chromosome number 2L followed by 2R, 3L, 3R, and X-chromosomes i.e. 2.10 +/- 0.44, 1.84 +/- 0.44, 1.57 +/- 0.54, 1.31 +/- 0.50, and 0.22 +/- 0.27 respectively. The susceptibility of different chromosomal arms to this pesticide was 2L > 2R > 3L > 3R > X and the regions prone to these aberrations have been marked on the polytene chromosome map of Anopheles maculatus.     

The chromosomes of Trypetesa lampas (Cirripedia,Acrothoracica)

Observations were made of the chromosomes of the burrowing barnacle Trypetesa lampas (Hancock). A method of squash preparation was used, which incorporated staining the material in a solution of 2% orcein in 45% acetic acid. The diploid number of chromosomes of T. lampas was found to be 12 and the haploid number 6, in both males and females. No obvious chromosomal mechanism of sex determination was found. There was, therefore, no cytogenetic confirmation of Kühnert's view (1934) that the sex of the larva is predetermined genetically. During mitosis in the females and embryos of Trypetesa lateralis Tomlinson and Kochlorine floridana Wells and Tomlinson, the diploid number of chromosomes was found to be about 14. The numbers of chromosomes in the 3 species of Acrothoracica studied were approximately half those observed in all 12 species of Thoracica and all 5 species of Rhizocephala previously investigated.     

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.     

Born to lose. I. Measures of tissue loss and regeneration by the brittlestar Microphiopholis gracillima (Echinodermata: Ophiuroidea)

To measure amounts of tissue lost in natural populations of the burrowing amphiurid ophiuroid Microphiopholis gracillima (Stimpson), individuals were collected from subtidal mud flats in North Inlet, South Carolina, USA, at monthly intervals between February 1985–February 1987 and December 1989–November 1990. Between 20 and 70% of all individuals were regenerating the disc, and 85% of the 2045 arms examined had regeneration scars; >50% had one scar and some arms had up to 4 scars. Fewer individuals were regenerating discs in warmer months, but there was no seasonality in arm-loss frequency. To quantify rates of arm regeneration in the field, individuals which had 1, 2, or 3 arms removed were placed in mud-filled cores in the field in late July and November 1988 and in March and May 1989, and recovered after periods of about one month. Another set of cores was held in a running seawater laboratory during the May 1989 experiment. No regeneration occurred during the cooler times of year (November and March), and rates of regeneration were slower in May (total: 0.13 mg/d; tissue: 0.03 mg/d) than July (total: 0.17 mg/d; tissue: 0.05 mg/d). These rates indicate complete replacement of lost tissue in 100 to 120 d during the growth season. Within experiments, per arm regeneration rates were similar regardless of the number of arms removed. This finding is complicated by small sample size, high variability and low statistical power, but in general individuals which lost 2 or 3 arms regenerated proportionally more tissue than individuals which lost 1 arm. Individuals held in the laboratory regenerated the same amount of tissue but 30% less skeleton than individuals in the field. Sublethal tissue loss is common in this population, and M. gracillima is capable of regenerating at least 50% (each arm=17% of total body weight x 3) of its standing crop in a single growing season. Burrowing brittlestars probably constitute a significant renewable energy source for higher trophic levels in areas where they occur in dense populations.     

Chromosomal nucleolar organizer region (NOR) phenotypes in nine species of the genus Ophryotrocha (Polychaeta: Dorvilleidae)

Chromosomal nucleolar organizer region (NOR) phenotypes have been characterized in nine species of the genus Ophryotrocha (Polychaeta: Dorvilleidae), namely O. notoglandulata, O. sp. macrovifera, O. sp. labronica pacifica, O. labronica labronica, O. puerilis puerilis, O. diadema, O. sp. robusta, O. gracills and O. hartmanni. Irrespective of chromosome number and morphology, Ag positive regions were terminally located in all but one species, O. diadema, where the NORs were pericentromerical in a metacentric pair. The presence of a single chromosome pair bearing NOR in invertebrates is considered an ancestral trait. According to this assumption, O. sp. robusta, O. dialema, and perhaps O. p. puerilis appear to be more ancestral than the other species. On the contrary, O. notoglandulaia, O. sp. macrovifera, O. sp. labronica oacifica, with two chromosomal pairs bearing NOR sites, seem to represent examples of further evolution within the genus Ophryotrocha.     

云南高黎贡山白颌大角蟾的核型,C—带及Ag—NORs的研究

比较了云南高黎贡山地区的贡山独龙江和腾冲大蒿坪白颌大角蟾（Megophryslateralis）两个地理种群的核型、C-带和Ag-NORs结果表明,两个地理种群在核型和带型上都有差异两个地理种群的核型均为2n＝26,NF＝52染色体形态差异不明显,而次缢痕的位置完全不同,贡山独龙江标本的次缢痕位于No．2的长臂上近着丝点处,腾冲标本的次缢痕位于No．5的短臂上近着丝点的部位在腾冲标本中发现一雄性个体中有一条额外的染色体,可能是B染色体两地标本的C-带差异不太显著,贡山独龙江标本的C-带相对较为显著．贡山独龙江标本的Ag-NORs位于No．2长臂近着丝点处,与次缢痕的部位对应,两条同源染色体上大小有显著差异．腾冲标本的Ag-NORs位于No．5短臂上近着丝点处,与次缢痕的部位对应依据核型和带型的比较,对白颌大角蟾的分类和进化问题进行了讨论。