Karyotypic analysis of the gobiid fish genus Quietula Jordan and Evermann

The karyotypes of the two species of the gobiid fish genus Quietula, Q. y-cauda (Jenkins & Evermann) and Q. guaymasiae (Jenkins & Evermann), are reported for the first time. The karyotypes contained equal numbers of chromosomes (2 n =42) but differed in chromosome morphology. Quietula y-cauda has 42 acrocentric chromosomes. Quietula guaymasiae has six metacentric, four submetacentric, and 32 acrocentric chromosomes. It is suggested the karyotype of Q. guaymasiae might have been derived from that of Q. y-cauda. In addition, from comparison of the species' karyotypes of the genera Quietula and Gillichthys , it is possible the karyotype of Q. y-cauda might have been derived from that of Gillichthys mirabilis.     

Karyotype analysis of Eucinostomus argenteus, E. gula, E. harengulus, and Eugerres plumieri (Teleostei, Gerreidae) from Florida and Puerto Rico

The karyotypes of four gerreids of the western Atlantic Ocean are documented. A diploid chromosome complement of 48 telocentric chromosomes was found in the four species (2N=48t, fundamental number FN=48). No differences were detected either in the number of chromosomes of the standard karyotype, in their karyotype size, or between the karyotypes derived from male or female specimens of any of the species. Chromosome length decreased progressively and slightly from pair 1 to pair 24. The Ag–NOR karyotypes of E. argenteus and E. harengulus were characterized by the position of the nucleolar organizer regions next to the centromere in chromosome pair 1, whereas in E. gula and E. plumieri Ag–NORs were detected in pair 4. The other 46 chromosomes showed a light staining of the centromere with no terminal or intermediate heterochromatic blocks. All Eucinostomus species showed Ag–NORs of similar size, while Eugerres plumieri showed Ag–NORs 10–20% larger than Eucinostomus species. A combination of size and position of the Ag–NORs identified E. gula, while size alone identified E. plumieri. However, the ancestral state for size and position of Ag–NORs could not be established. There was no differential staining of the chromosomes by G-banding. The karyotype of the gerreids appears similar to the hypothetical ancestral karyotype of fish. The phylogenetic relationships among these species could not be established because of the lack of chromosome G-bands. Most likely this indicates a homogeneous distribution of GC nucleotides in the chromosomes.     

More sex chromosomes than autosomes in the Amazonian frog <Emphasis Type="Italic">Leptodactylus pentadactylus</Emphasis>

Heteromorphic sex chromosomes are common in eukaryotes and largely ubiquitous in birds and mammals. The largest number of multiple sex chromosomes in vertebrates known today is found in the monotreme platypus (Ornithorhynchus anatinus, 2n?=?52) which exhibits precisely 10 sex chromosomes. Interestingly, fish, amphibians, and reptiles have sex determination mechanisms that do or do not involve morphologically differentiated sex chromosomes. Relatively few amphibian species carry heteromorphic sex chromosomes, and when present, they are frequently represented by only one pair, either XX:XY or ZZ:ZW types. Here, in contrast, with several evidences, from classical and molecular cytogenetic analyses, we found 12 sex chromosomes in a Brazilian population of the smoky jungle frog, designated as Leptodactylus pentadactylus Laurenti, 1768 (Leptodactylinae), which has a karyotype with 2n?=?22 chromosomes. Males exhibited an astonishing stable ring-shaped meiotic chain composed of six X and six Y chromosomes. The number of sex chromosomes is larger than the number of autosomes found, and these data represent the largest number of multiple sex chromosomes ever found among vertebrate species. Additionally, sequence and karyotype variation data suggest that this species may represent a complex of species, in which the chromosomal rearrangements may possibly have played an important role in the evolution process.     

A chromosome study of Spanish Bembidiidae and other Caraboidea (Coleoptera,Adephaga)

The karyotypes were studied of fifty-three Spanish Caraboidea species belonging to the following families: Trechidae (3 species), Bembidiidae (48), Pogonidae (1) and Harpalidae (1). Differences in the shape of the largest pair of autosomes (A pair) and the Y-chromosome are frequent among closely related species. The chromosomes of these species have a peculiar centromeric region weakly constricted and surrounded by conspicuous heterochromatic blocks. Eight species have a heteromorphic A pair, probably due to differences in the length of the nucleolar organizer. Species of the first three families either have a male karyotype of 24 chromosomes, or one derived from it, while the only species of the fourth family has a male karyotype of 39 chromosomes derived from the basic karyotype of the Harpalidae, 2n=37.     

B chromosomes in two fish species, genus Rhamdia (Siluriformes, Pimelodidae)

Specimens belonging to two fish species genus Rhamdia were cytogenetically analysed from seven localities in Brazil and Argentina. In addition to the 58 chromosomes of the basic karyotype, one to five metacentric B chromosomes were observed carrying conspicuous heterochromatic blocks on the distal regions of both chromosome arms. These B chromosomes are mitotically stable and, in the two best-sampled populations in R. hilarii (Lobo and 29 reservoirs), they showed frequency distributions fitting a binomial distribution, though Bs were more frequent in the latter. The presence of B chromosomes with the same appearance in R. quelen suggests an ancient origin for these B chromosomes, presumably prior to speciation from a common ancestor.     

Karyotype differentiation between two stickleback species (Gasterosteidae)

The stickleback family (Gasterosteidae) of fish is less than 40 million years old, yet stickleback species have diverged in both diploid chromosome number (2n) and morphology. We used comparative fluorescence in situ hybridization (FISH) on 2 stickleback species, Gasterosteus aculeatus (2n = 42) and Apeltes quadracus (2n = 46), to ascertain the types of chromosome rearrangements that differentiate these species. The A. quadracus karyotype contains more acrocentric and telocentric chromosomes than the G. aculeatus karyotype. By using bacterial artificial chromosome probes from G. aculeatus in our FISH screen, we found that 6 pericentric inversions and 2 chromosome fusions/fissions are responsible for the greater number of acrocentric and telocentric chromosomes in A. quadracus. While most populations of G. aculeatus have an XX/XY sex chromosome system, A. quadracus has a ZZ/ZW sex chromosome system, as previously reported. However, we discovered that a population of A. quadracus from Connecticut lacks heteromorphic sex chromosomes, providing evidence for unexpected sex chromosome diversity in this species.     

Karyotype diversity of four species of the incertae sedis group (Characidae) from different hydrographic basins: analysis of AgNORs, CMA3 and 18S rDNA

A large number of genera in the tropical fish family Characidae are incertae sedis. Cytogenetic analysis was made of four of these species: Astyanax eigenmanniorum, Deuterodon stigmaturus, Hyphessobrycon luetkenii, and H. anisitsi, collected from various hydrographic basins: hydrographic system from Laguna dos Patos/RS, Tramandaí basin/RS and Tibagi River basin/PR. The first two species were collected in their type locality in the State of Rio Grande do Sul. The 2n = 48 karyotype was observed only in A. eigenmanniorum, while the other species had 2n = 50 chromosomes, with different karyotypic formulas. There was weak heterochromatin staining in the pericentromeric region of A. eigenmanniorum, D. stigmaturus and H. luetkenni chromosomes. In H. anisitsi, heterochromatin appeared to be more abundant and distributed in the pericentromeric and terminal regions of the chromosomes; three pairs showed more evident heterochromatic blocks. There were multiple Ag-NORs in all populations, visualized by FISH with an 18S rDNA probe. While D. stigmaturus and H. luetkenii had conserved AgNOR, CMA3 and 18S rDNA sites, the other two species showed intra- and interindividual variation at these sites. The karyotype variability was high, as is common in this group of fish. Different species arising from isolated hydrographic basins maintain an elevated level of karyotype differentiation, mainly with respect to chromosome structure, heterochromatin distribution and rDNA localization. This is the first report with cytogenetic data for D. stigmaturus and H. luetkenii.     

Sex chromosomes, sex determination, and sex-linked sequences in Microtidae

The Arvicolidae is a widely distributed rodent group with several interesting characteristics in their sex chromosomes. Here, we summarize the actual knowledge of some of these characteristics. This mammalian group has species with abnormal sex determination systems. In fact, some species present the same karyotype in both males and females, with total absence of a Y chromosome, and hence of SRY and ZFY genes. Other species present fertile, sex-reversed XY females, generally due to mutations affecting X chromosomes. Furthermore, in Microtus oregoni males and females are gonosomic mosaic (the females are XO in the soma and XX in the germ cells, while the males are XY in the soma and OY in the germ cells). Regarding sex chromosomes, some species present enlarged (giant) sex chromosomes because of the presence of large blocks of constitutive heterochromatin, which have been demonstrated to be highly heterogeneous. Furthermore, we also consider the alterations affecting composition and localization of sex-linked genes or repeated sequences. Finally, this rodent group includes species with synaptic and asynaptic sex chromosomes. In fact, several species with asynaptic sex chromosomes have been described. It is interesting to note that within the genus Microtus both types of sex chromosomes are present.     

Chromosome complement, C-banding and Ag-NOR in Gymnammodytes cicerelus (Ammodytidae, Perciformes)

The karyotype of a sandlance species, Gymnammodytes cicerelus , comprises: seven meta-centric, seven submetacentric and nine subtelocentric-acrocentric pairs (2 n =46, FN=74). The C-bands appear in paracentromeric and telomeric areas of most chromosomes and the NOR regions, in two pairs of larger chromosomes. All these characteristics indicate that a large number of rearrangements seem to have been involved in the karyotype evolution of this species.     

Karyotype and genome size of zoarcids and notothenioids (Taleostei,Perciformes) from the Ross Sea: cytotaxonomic implications

In the absence of fossils, the origin of Notothenioidei, a perciform suborder dominating the fish fauna of the Southern Ocean, remains conjectural; some morphoecological evidence suggests relationships to zoarcoids. To test this point we have compared the karyotype morphology and genome size of two species of zoarcids from the Ross Sea to those of one species each of the notothenioid families Artedidraconidae, Bathydraconidae, Channichthyidae and Nototheniidae from the same region. A karyotype of 48, mostly acrocentric, chromosomes, localization of nucleolar organizers on a pair of small dibrachial chromosomes, a genome size of about 3 pg of DNA, characterize both zoarcids; similar features can be found in the karyology of the notothenioids (especially the Nototheniidae). However, all shared characters appear as plesiomorphic in teleost karyology, which does not help in producing new data on the problem of notothenioid relationships.     

Assessing Genetic Diversity of Brazilian Reef Fishes by Chromosomal and DNA Markers

Little is known on genetics of Brazilian coral reef fish and most of this information is limited to chromosome characterization of major representative species. The diploid chromosome number in marine fish varies from 2n= 22–26 to 2n = 240–260. Despite of this apparent diversity, most studied marine species have a diploid complement with 48 acrocentric chromosomes. This latter trend is mostly observed among Perciformes, an important major taxon of coral reef fishes. Studies in the families Pomacentridae, Pomacanthidae and Chaetodontidae, for example, have shown a common karyotype pattern entirely formed by 48 uniarmed chromosomes. However, rare numerical and structural chromosome polymorphisms and cryptic chromosome rearrangements involving heterochromatin segments and/or nucleolar organizing sites have been reported among such fishes. Although new chromosome forms can contribute to the establishment of genetically isolated populations, their role in reef fish speciation at marine realm still is an open question. More recently, genomic DNA analyses using RAPD and microsatellites, and sequencing and RFLP of mitochondrial DNA have increasingly been used in Atlantic reef fish species. Genetic homogeneity over wide geographical ranges has been reported for different fish groups, in contrast to several cases of population substructuring related to environmental constraints or evolutionary history. Amazonas outflow and upwelling on the Southeastern coast of Brazil are believed to be strong barriers to dispersal of some reef species. Moreover, it is suggested that the pattern of speciation and population structure at South Atlantic is quite distinctive from Pacific Ocean, even when comparing closely related taxa. Further genetic studies are strongly encouraged in Brazilian reef fishes in order to provide a reliable scenario of the genetic structure in this important and diverse fish group.     

尼罗尖吻鲈和鳜鱼染色体组型分析及比较

采用PHA、秋水仙碱腹腔或背部肌肉注射,活体培养法,以前肾为材料,低渗-空气干燥法制片,进行染色体观察,运用Micromeasure version 3.3染色体分析软件和Photoshop 7.0软件首次分析了尼罗尖吻鲈的染色体数目和核型,并同鳜鱼染色体数目和核型进行了分析比较,对今后拟采取的杂交尝试提供理论基础。结果显示：尼罗尖吻鲈染色体众数为2n=48,核型公式为2m+4sm+12st+30t,染色体臂数（NF）为54;鳜鱼染色体众数为2n=48,核型公式为6sm+12st+30t,染色体臂数（NF）亦为54;两种鱼染色体短臂上均无随体,单臂染色体较多。分析表明尼罗尖吻鲈与鳜鱼杂交成功的可能性较大。     

Molecular and cytogenetic analysis of the telomeric (TTAGGG)n repetitive sequences in the Nile tilapia,Oreochromis niloticus (Teleostei: Cichlidae)

The majority of chromosomes in Oreochromis niloticus, as with most fish karyotyped to date, cannot be individually identified owing to their small size. As a first step in establishing a physical map for this important aquaculture species of tilapia we have analyzed the location of the vertebrate telomeric repeat sequence, (TTAGGG)n, in O. niloticus. Southern blot hybridization analysis and a Bal31 sensitivity assay confirm that the vertebrate telomeric repeat is indeed present at O. niloticus chromosomal ends with repeat tracts extending for 4-10 kb on chromosomal ends in erythrocytes. Fluorescent in situ hybridization revealed that (TTAGGG)n is found not only at telomeres, but also at two interstitial loci on chromosome 1. These data support the hypothesis that chromosome 1, which is significantly larger than all the other chromosomes in the karyotype, was produced by the fusion of three chromosomes and explain the overall reduction of chromosomal number from the ancestral teleost karyotype of 2n=48 to 2n=44 observed in tilapia.     

C-banded chromosomes of the genus Bembidion Latr. (Coleoptera, Carabidae)

The C-banding pattern of Bembidion punctulatum, B. varium, B. varicolor, B. ascedens, B. tibiale, B. ustulatum, B. decorum, and B. modestum are presented. All examined species have a symmetrical karyotype with meta- and submentacentric chromosomes and meioformula n=11+XY. All of them have an achiasmatic spermatogenesis in common. The present data confirm earlier studies indicating a considerable morphological and numerical stability of the Bembidiini karyotype. The C-banding showed the existence of heterochromatin in the paracentric regions of chromosomes, and also 2-3 intercalar C-positive segments were observed. The Y chromosome is entrely euchromatic. The C-banded karyotype of the analysed species in genus Bembidion has large heterochromatin segments on chromosomes, an exception in Coleoptera.     

The discovery of triploid Lycoris sprengeri Comes ex Baker from Anhui,China

Lycoris sprengeri Comes ex Baker is endemic to China. Reported in the present paper are the chromosomes number and karyotypes for two wild populations of the species from Anhui. ( 1 )Caishi population has a karyotype 2n=33=9st+21t+3T. The length of chromosomes ranges from 5.58～9.15μm. The karyotype belongs to Stebbin’s (1971) “4A”. (2)Longyashan populations have two karyotypes. The karyotype formula of the type I is 2n=22=8st+14t, with chromosomes ranging from 6.88～9.15μm. The karyotype belongs to “4A”. The karyotype formula of the type Ⅱ is 2n＝22＝1m+1sm+14st+6t, with chromosomes ranging from 7.20～15.80μm. The karyotype belongs to “3B”. The triploid type of L. sprengeri was discovered in Anhui for the first time. The karyotype 2n=22 =1m+1sm+14st+6t in diploid type of this species is here reported for the first time.The Robertsonian change plays a key role in karyotype evolution of Lycoris.     

Karyotype and mitochondrial 16S gene characterizations in seven South American Cichlasomatini species (Perciformes, Cichlidae)

The family Cichlidae constitutes most of the freshwater fish fauna of South America; its taxonomy is at present mainly based on morphological characters. Here, relationships among seven Cichlasomatini species have been investigated by studying their karyotype structure and by sequencing a 520 bp fragment of the mitochondrial 16S gene. Molecular data sets point to a high affinity of Cichlasoma amazonarum with Aequidens sensu stricto group, in particular with Aequidens tetramerus . Aequidens never form a single monophyletic clade: molecular trees group together ' Aequidens pulcher ' and ' Aequidens rivulatus ' and suggest their close relationship with Bujurquina and Laetacara , rather than with the A. sensu stricto group. Both molecular and karyotypic data confirm that Cleithracara maronii belongs to a distinct clade, thus supporting its generic differentiation based on morphological characters. Chromosome number, karyotype structure and molecular data suggest that Laetacara dorsigera is related to Bujurquina vittata and confirm their generic level of differentiation. From a cytotaxonomic point of view, a karyotype of 2 n  = 48 with most acrocentric or subacrocentric chromosomes could be the ancestral one from which the others might have derived.     

Cytogenetic analysis of a hybrid, Glyptotendipes pallens Mg. × Glyptotendipes glaucus Mg. (Diptera, Chironomidae): evolutionary considerations

The karyotype of experimentally obtained hybrids between the two closely related species Glyptotendipes pallens and Glyptotendipes glaucus is described. Hybridization was successful in one direction only ( G. pallens ♂ x G. glaucus ♀). The polytene chromosomes AB and EF of the hybrid show a more or less intimate pairing throughout their length. In the chromosomes CD in which an inversion occurs the characteristic loop is formed. The homologues of chromosome G are almost completely asynaptic. The localization of centromere heterochromatin was also studied. Centromere heterochromatin as well as intercalary heterochromatin could be observed in all chromosomes. By C banding analyses it could be shown that G. pallens has a telomeric chromosome G while in G. glaucus it is acrocentric. According to karyotype similarity it can be assumed that these two species have quite recently derived from a common ancestor since they still share much of their genomic organization. On the Black Sea coast (southeast part of Bulgaria) a natural hybridization zone between the sympatric species G. pallens and G. glaucus has been detected. The idea that hybridization between the two species might finally proceed to the formation of a new species by hybrid origin and introgression is discussed.     

Highly differentiated and conserved sex chromosome in fish species (Aulopus japonicus: Teleostei, Aulopidae)

While highly differentiated and long-conserved sex chromosomes such as XY and ZW chromosomes are observed, respectively, in mammalian and avian species, no counterparts to such chromosomes were observed in fish until we reported in the previous study that well-conserved and highly differentiated ZW sex chromosomes existed in the family of Synodontidae. Then, the problem was if the evolutionary history of the fish ZW chromosomes was long enough to be comparable to the mammalian and avian counterparts. To tackle the problem, we had to extend our finding of the fish sex chromosomes further than a family alone. For this purpose, we chose Aulopus japonicus that belonged to one of the related families to Synodontidae.Our cytogenetic and fluorescence in situ hybridization (FISH) analyses have clearly demonstrated that A. japonicus also has ZW chromosomes. We have also found that 5S rDNA clusters are located on the Z and W chromosomes in this species. Using nontranscribed intergenic sequences in the 5S rDNA clusters as PCR primers, we successfully amplified a 6-kb-long female-specific sequence on the W chromosome. The 6-kb-long sequence contained one transposable element and two tRNA sequences. The function of the sequence remains to be studied. Our Southern blot analysis confirmed that the 6-kb sequence was located only on the W chromosome.Therefore, it is now said that highly differentiated ZW chromosomes have been conserved over two fish families. As these families were reported to have been diverged 30-60 million years ago, the fish ZW chromosomes have an evolutionary history corresponding to the history of the families. This is perhaps the first case that fish sex chromosomes are shown to have such a long evolutionary lineage.     

Evidence for chromosome number reduction and chromosomal homosequentiality in the 24-chromosome Korean frog Rana dybowskii and related species

The karyotype of the Korean frog Rana dybowskii with its pattern of C-band heterochromatin distribution was numerically analyzed. There are 2n = 24 chromosomes in the karyotype representing a reduction in number from the typical 2n = 26 chromosome karyotype of Rana. The karyotype shows other evidence of reorganization relative to 26-chromosome species. The chromosomes grade smoothly in size from largest to smallest without the two size classes that are characteristic for 26-chromosome species. In contrast to many 26-chromosome species, there are few centromeric C-bands but many interstitial ones. C-bands for each homologous chromosome pair are distinctive. A prominent secondary constriction is located on one of the smallest chromosomes, chromosome 11, in a position similar to that seen in most 26-chromosome species. The karyotype of R. dybowskii is compared to those of other species of Rana known to have 2n = 24 chromosomes; it is most similar to that of R. chensinensis, less so that of R. ornativentris and less still to that of R. arvalis in terms of the positions of centromeres and secondary constrictions. C-bands as well as secondary constrictions in the karyotypes of these frogs show evidence of chromosomal homosequentiality. The process and possible consequences of chromosome number reduction from an ancestral 26-chromosome karyotype is also evident in the karyotypes of these closely allied palearctic frogs. Pericentric inversions followed by fusion of two small elements apparently produced a new chromosome, chromosome 6, occurring originally among northeast Asian populations.