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.     

Cytogenetic characterization and mapping of rDNAs,core histone genes and telomeric sequences in <Emphasis Type="Italic">Venerupis aurea</Emphasis> and <Emphasis Type="Italic">Tapes rhomboides</Emphasis> (Bivalvia: Veneridae)

We describe the chromosomal location of GC-rich regions, 28S and 5S rDNA, core histone genes, and telomeric sequences in the veneroid bivalve species Venerupis aurea and Tapes (Venerupis) rhomboides, using fluorochrome staining with propidium iodide, DAPI and chromomycin A3 (CMA) and fluorescent in situ hybridization (FISH). DAPI dull/CMA bright bands were coincident with the chromosomal location of 28S rDNA in both species. The major rDNA was interstitially clustered at a single locus on the short arms of the metacentric chromosome pair 5 in V. aurea, whereas in T. rhomboides it was subtelomerically clustered on the long arms of the subtelocentric chromosome pair 17. 5S rDNA also was a single subtelomeric cluster on the long arms of subtelocentric pair 17 in V. aurea and on the short arms of the metacentric pair 9 in T. rhomboides. Furthermore, V. aurea showed four telomeric histone gene clusters on three metacentric pairs, at both ends of chromosome 2 and on the long arms of chromosomes 3 and 8, whereas histone genes in T. rhomboides clustered interstitially on the long arms of the metacentric pair 5 and proximally on the long arms of the subtelocentric pair 12. Double and triple FISH experiments demonstrated that rDNA and H3 histone genes localized on different chromosome pairs in the two clam species. Telomeric signals were found at both ends of every single chromosome in both species. Chromosomal location of these three gene families in two species of Veneridae provides a clue to karyotype evolution in this commercially important bivalve family.     

菠菜rDNA及端粒多色荧光原位杂交分析

以拟南芥25S rDNA、5S rDNA以及端粒序列为探针对菠菜的中期染色体进行了多色荧光原位杂交分析, 其中25S rDNA用生物素标记, 端粒序列用地高辛标记, 5S rDNA用生物素和地高辛共同标记。杂交结果显示, 菠菜中期染色体25S rDNA杂交位点为6个, 分别位于3号、5号和6号染色体的随体部位; 5S rDNA杂交位点为4个, 分别位于3号和5号染色体长臂, 端粒序列杂交位点位于6号染色体端部以及其余染色体的端部和着丝粒部位。     

Major cytogenetic landmarks and karyotype analysis inDaucus carota and other Apiaceae

Karyotype analysis provides insights into genome organization at the chromosome level and into chromosome evolution. Chromosomes were marked for comparative karyotype analysis using FISH localization of rDNA genes for the first time in Apioideae species including taxa of economic importance and several wild Daucus relatives. Interestingly, Daucus species did not vary in number of rDNA loci despite variation in chromosome number (2n = 18, 20, 22, and 44) and previous publications suggesting multiple loci. All had single loci for both 5S and 18S-25S (nucleolar organizing region) rDNA, located on two different chromosome pairs. The 5S rDNA was on the short arm of a metacentric chromosome pair in D. crinitus (2n = 22) and D. glochidiatus (2n = 44) and on the long arm of a metacentric pair in other Daucus species, suggesting possible rearrangement of this chromosome. For other Apiaceae, from two (Apium graveolens), to three (Orlaya grandiflora), to four (Cuminum cyminum) chromosomes had 18S-25S rDNA sites. Variability for number and position of the 5S rDNA was also observed. FISH signals enabled us to identify 20-40% of the chromosome complement among species examined. Comparative karyotype analysis provides insights into the fundamental aspects of chromosome evolution in Daucus.     

Molecular cytogenetics and characterization of a ZZ/ZW sex chromosome system in Triportheus nematurus (Characiformes, Characidae)

Chromosomes of Triportheus nematurus, a fish species from family Characidae, were analyzed in order to establish the conventional karyotype, location of C-band positive heterochromatin, Ag-NORs, GC- and AT-rich sites, and mapping of 18S and 5S rDNA with fluorescence in situ hybridization (FISH). The diploid number found was 2n = 52 chromosomes in both males and females. However, the females presented a pair of differentiated heteromorphic chromosomes, characterizing a ZZ/ZW sex chromosome system. The Z chromosome was metacentric and the largest one in the karyotype, bearing C-positive heterochromatin at pericentromeric and telomeric regions. The W chromosome was middle-sized submetacentric, appearing mostly heterochromatic after C-banding and presenting heterogeneous heterochromatin composed of GC- and AT-rich regions revealed by fluorochrome staining. Ag-NORs were also GC-rich and surrounded by heterochromatic regions, being located at the secondary constriction on the short arms of the second chromosome pair, in agreement with 18S rDNA sites detected with FISH. The 18S and 5S rDNA were aligned in tandem, representing an uncommon situation in fishes. The results obtained reinforce the basal condition of the ZZ/ZW sex system in the genus Triportheus, probably arisen prior to speciation in the group.     

Classical and molecular cytogenetics and DNA content in Maihuenia and Pereskia (Cactaceae)

We studied cacti species of the subfamilies Pereskioideae (five species of the southern clade) and both species of Maihuenioideae using molecular cytogenetic techniques and DNA content. Mitotic chromosomes were analyzed for Pereskia aculeata, P. bahiensis, P. grandifolia, P. nemorosa, P. sacharosa, Maihuenia poeppigii, and M. patagonica, using the Feulgen stain, CMA/DAPI fluorescent chromosome banding, fluorescence in situ hybridization (FISH, probes of 5S rDNA and pTa71 for 18-5.8-26S rDNA), and DNA content by flow cytometry technique. The karyotypes were highly symmetrical, most of the pairs being metacentric (m). CMA/DAPI banding revealed the presence of CMA⁺/DAPI^? bands associated with NORs in the first m pair of all species. The co-localization of 18-5.8-26S rDNA loci with CMA⁺/DAPI^?/NORs blocks allowed the identification of homeologous chromosome pairs between species of both subfamilies. FISH using probe 5S rDNA was applied for the first time in both subfamilies. Diploid species had always one m pair carrying 5S rDNA genes, with pericentromeric location in different chromosome pairs. In the tetraploid cytotype of M. patagonica, the 5S rDNA probe hybridized to two pairs. The 2C DNA content obtained by FC varied twofold (from 1.85 to 2.52 pg), with significant differences between species. Mean chromosome length, karyotype formula, percentage of heterochromatin position of 5S rDNA locus, and nuclear Cx DNA content vary among Maihuenia and Pereskia species and allowed to differentiate them. Both genera are closely related and that the differences found are not strong enough to separate Maihuenioideae from Pereskioideae.     

慈姑45S rDNA和端粒序列检测及核型分析

以45S r DNA和拟南芥型端粒序列为探针对慈姑(Sagittaria trifolia L.)有丝分裂中期染色体进行单色和双色荧光原位杂交分析,并用银染方法检测慈姑45S r DNA位点的表达,最后结合染色体测量数据和45S r DNA杂交信号建立慈姑的核型。结果显示,慈姑的单倍基因组总长度为76.9±1.38μm,最长染色体为11.55±0.10μm,最短染色体为4.54±0.27μm;慈姑的核型公式为:2n=22=2m+2sm+14st+4t,核型不对称性参数CI、A1、A2、As K(%)、AI分别为19.86±11.06、0.72、0.27、78.82、15.29,核型属于Stebbins类型中的3B型。慈姑具有3对45S r DNA位点,分别位于第8、9、10号染色体的短臂末端。拟南芥型端粒序列的杂交信号出现在慈姑每一条染色体的长、短臂末端。银染检测到6个Ag-NOR和6个核仁,表明3对45S r DNA位点在间期核都有表达。本研究结果为药食兼用植物慈姑提供了分子细胞遗传学基础资料。     

Comparative molecular cytogenetic analysis of three <Emphasis Type="Italic">Leuciscus</Emphasis> species (Pisces,Cyprinidae) using chromosome banding and FISH with rDNA

A comparative molecular cytogenetic analysis was performed on three species of the genus Leuciscus viz. ide L. idus, chub L. cephalus and dace L. leuciscus distributed in Poland, using C-, Ag- and chromomycin A₃ (CMA₃)-stainings and fluorescence in situ hybridization (FISH) with 5.8S + 28S rDNA as a probe. Although the three species examined shared 2n = 50 chromosomes and the largest acrocentric chromosome pair in the complement, they were characterized with karyotypic differences in terms of the number of uni- and biarmed chromosomes and the localization of nucleolar organizer regions (NORs) revealed by Ag-staining and FISH. L. idus and L. cephalus showed the rDNA sites on the long arms of one submetacentric (SM) chromosome pair and on the short arms of one subtelocentric (ST) chromosome pair, respectively. These NORs were CMA₃-positive, GC-rich and C-positive heterochromatic sites in both species. Such chromosome banding features were also true for four NORs localizing on one of each SM and ST pair in L. leuciscus, but considerable numerical NOR polymorphism became apparent with Ag-staining and FISH due to a different combination of these NOR-bearing SMs and STs in this dace. The present results indicate that the molecular cytogenetic analysis applied herein may become useful to elucidate the karyotype evolution and phylogenetic relationships among the species in the genus Leuciscus and other related groups.     

Chromosomal localization of 45S and 5S rDNA in 14 species and the implications for genome evolution of genus Epimedium

Studying the genome structure of Epimedium has been hindered by the large genomes and uniform karyotypes. Consequently our understanding of the genome organization and evolutionary changes of Epimedium is extremely limited. In the present study, the 45S and 5S rDNA loci of 14 Epimedium species were physically mapped by double-probe FISH for the first time. Results showed the following: (1) Chromosomes I and II of all 14 species examined, except for E. shuichengense, hosted one pair of 45S rDNA sites, respectively. Most of the 45S rDNA sites gave clear signals and were positioned in the distal regions of the short arms. (2) All species studied of section Diphyllon were found to have one pair of 5S rDNA sites localized in the interstitial regions of the long arm of chromosome IV, and the two species of section Epimedium, E. alpinum and E. pubigerum, had two pairs of 5S rDNA sites localized in the interstitial regions of the long arm of chromosomes IV and V, respectively. (3) In section Diphyllon, all species of small flower taxa, except E. shuichengense, had three pairs of 45S rDNA sites, clearly more than species of big flower taxa, except E. davidii, with two pairs of 45S rDNA sites. Based on the 45S and 5S rDNA distribution patterns and other chromosomal morphological characteristics, six pairs of chromosomes can be unambiguously identified in all 14 Epimedium species. The stable differentiation in 45S and 5S rDNA FISH patterns between the two sections suggests that chromosomal rearrangements and transpositional events played a role in the splitting of the two sections, and section Diphyllon may be more primitive than section Epimedium. In the same way, big flower taxa may be more primitive than small flower taxa in section Diphyllon.     

Chromosome analysis of walking catfish,Clarias magur (Teleostei,Siluriformes, Clariidae), using staining,FISH with 18S, 5S and BAC probes

The chromosomal characteristics of Clarias magur were examined using conventional (Giemsa-staining, Ag-impregnation and CMA₃ + DAPI fluorescence) and molecular/ FISH (18S & 5S rDNA probes + one BAC DNA probe) cytogenetic tools. The diploid chromosome number was 50 and the karyotype consisted of 14 metacentric, 20 sub-metacentric, 8 sub-telocentric, 8 acrocentric chromosomes with 84 chromosome arms without any heteromorphic pair. The C-heterochromatic blocks were located on centromeric position of 13 pairs of chromosomes. The NOR sites, visualized by AgNO₃- and CMA₃- staining, were situated at p arms of chromosome pair No. 21, which also corresponded to 18S rDNA site visualized by FISH. The FISH signal of ICF_001_D19 clone probe was observed on 18th chromosome pair. The findings of the present study on C. magur provided valuable markers for the chromosome identification and locations of genes of the BAC clone on the chromosome will lead to the construction of physical map of genome of this species.     

Diapausing egg banks,lake size,and genetic diversity in the rotifer <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> Müller (Rotifera,Monogononta)

The fish from the family Anostomidae represent one of the most important groups of freshwater ichthyofauna from South America, with species of high economical value. The migratory characteristic of some species, through the several Amazonian environments, takes them into waters with different physico-chemical characteristics. Cytogenetic studies on the Anostomidae demonstrate that these fishes have a conserved diploid number and karyotype macrostructure. So, to verify if this conservation occurs also in the genomic level, the current study aimed at a chromosomal comparative physical mapping, using 45S and 5S rDNA, of seven species of anostomids: Leporinus fasciatus, L. agassizi, L. friderici, L. trifasciatus, Rhytiodus macrolepis, Laemolyta taeniata, and Schizodon fasciatus, collected in different Amazonian environments. The results obtained corroborate the conservation of the karyotype macrostructure. However, significant differences were found in the distribution of heterochromatin and on the pair bearing the nucleolus organizer region. The staining of 45S and 5S rDNA by FISH highlighted, for four of the seven species, more than one chromosome pair bearing the site 45S. The 5S rDNA, although present in only one chromosome pair, varied in its chromosome and karyotype position. Thus, although the Anostomidae family has a conserved chromosomic macrostructure the use of molecular techniques revealed the presence of chromosomic translocation during the evolution of these fishes.     

45S rDNA和5S rDNA在南瓜、丝瓜和冬瓜染色体上的比较定位

首次利用荧光原位杂交和双色荧光原位杂交技术对45S和5S rDNA在南瓜(Cucurbita moschata Duch)、丝瓜(Luffa cylindrical Roem)、冬瓜(Benincasa hispida Cogn)的有丝分裂中期染色体上进行了物理定位分析。南瓜有5对45S rDNA位点, 2对5S rDNA位点; 丝瓜具有5对45S rDNA位点, 1对5S rDNA位点; 冬瓜具有2对45S rDNA位点, 1对5S rDNA位点, 5S rDNA位点与其中一对45S rDNA位点都位于7号染色体短臂上, 并在物理位置上紧密相邻。45S rDNA在这3种作物染色体上数目变化较大, 但在染色体上都倾向分布在短臂末端, 其分布模式较为一致。5S rDNA在这3种作物染色体上数目相对保守, 但在染色体上分布的位置变化较大。文中讨论了45S rDNA和5S rDNA在植物基因组中不同的进化趋势。     

Karyotyping of three Pinaceae species via fluorescent in situ hybridization and computer-aided chromosome analysis

The positions of 18/25S rRNA genes, 5S RNA genes and of Arabidopsis-type telomeric repeats were localized by fluorescent in situ hybridization (FISH) on the chromosomes of three coniferous species; Picea abies, Larix decidua and Pinus sylvestris, each with 2n=24 chromosomes. Computer-aided chromosome analysis was performed on the basis of the chromosome length, the arm length ratio and the position of the hybridization signals. This enabled the chromosomes of the Norway spruce, 4 chromosomes of the European larch and 3 of the karyotype of the Scots pine to be individually distinguished. With respect to the chromosomal positions of rDNA and 5S rDNA loci, chromosome pair I of P. sylvestris is suggested to be homoeologous to pair II of P. abies, while another chromosome pair of P. sylvestris might be homoeologous to chromosome pair III of L. decidua.     

Characterization of Japanese flounder karyotype by chromosome bandings and fluorescence in situ hybridization with DNA markers

The chromosomes of Japanese flounder, Paralichthys olivaceus, were examined by conventional differential staining methods including G-, Q-, C-, silver (Ag)-, fluorochrome, and replication R-bandings and by fluorescence in situ hybridization (FISH) with 5S and 18S rDNAs and telomeric DNA as probes. Replication R-banding substantially made it possible to identify 24 homologous pairs by their RBG-banding pattern and relative length. Both rDNA loci were mapped to chromosome 1, where 5S and 18S rDNA loci were located at the centromeric region and secondary constriction, respectively. C-banding revealed that both rDNA loci were heterochromatic, and 18S rDNA loci were positive for chromomycin A₃ but negative for 4′,6-diamidino-2-phenylindole (DAPI) staining. Telomeric FISH signals were observed at all chromosome ends and at the interstitial region of some chromosomes. The observed results were discussed in relation to the karyotype evolution in the order Pleuronectiformes.     

基于荧光原位杂交的藜属植物核型分析

为精确地识别藜属植物染色体组的核型特征,该文研究了4种来自青海高原的野生藜属植物(灰绿藜、藜、菊叶香藜及杂配藜)和1种从美国引进的栽培藜麦品种PI614932-HX(3)基于染色体荧光原位杂交(rDNA FISH)的核型。利用5S rDNA和45S rDNA对5种藜属植物有丝分裂中期的染色体进行FISH研究。藜属植物的核型分析结果表明:(1)藜属植物中存在二倍体(2n=2x=18)和四倍体(2n=4x=36)两种倍性,藜麦和灰绿藜为四倍体,其余3种为二倍体。(2)藜麦、灰绿藜、藜、菊叶香藜及杂配藜的核型公式分别为2n=4x=36=34m(2AST)+2sm,2n=4x=36=32m(4AST)+4sm,2n=2x=18=16m(4AST)+2sm,2n=2x=18=18m及2n=2x=18=16m+2sm。(3)染色体由大部分的中部着丝粒染色体(m)和少部分近中部着丝粒染色体(sm)组成。(4)核型类型除了菊叶香藜为1B以外,其余均属于2B类型。(5)在藜麦、灰绿藜及藜中具有分布位置不同、数量不等的双随体。5S rDNA、45S rDNA FISH结果表明:(1)藜麦和灰绿藜的染色体上存在2对5S rDNA位点和1对45S rDNA位点,藜、杂配藜的染色体上存在1对5S rDNA位点和1对45S rDNA位点,菊叶香藜的染色体上只存在1对5S rDNA位点。(2)5S rDNA和45S rDNA位点均位于染色体的短臂上。该研究首次获得了藜属植物基于5S rDNA和45S rDNA荧光原位杂交核型,为藜属植物亲缘关系研究和细胞生物学研究提供了分子细胞遗传学依据。     

Relationships of the woody Medicago species (section Dendrotelis) assessed by molecular cytogenetic analyses

Background and Aims

The organization of rDNA genes in the woody medic species from the agronomically important Medicago section Dendrotelis was analysed to gain insight into their taxonomic relationships, to assess the levels of infraspecific variation concerning ribosomal loci in a restricted and fragmented insular species (M. citrina) and to assess the nature of its polyploidy.

Methods

Fluorescence in situ hybridization (FISH) was used for physical mapping of 5S and 45S ribosomal DNA genes in the three species of section Dendrotelis (M. arborea, M. citrina, M. strasseri) and the related M. marina from section Medicago. Genomic in situ hybridization (GISH) was used to assess the genomic relationships of the polyploid M. citrina with the putatively related species from section Dendrotelis.

Key Results

The diploid (2n = 16) M. marina has a single 45S and two 5S rDNA loci, a pattern usually detected in previous studies of Medicago diploid species. However, polyploid species from section Dendrotelis depart from expectations. The tetraploid species (2n = 32) M. arborea and M. strasseri have one 45S rDNA locus and two 5S rDNA loci, whereas in the hexaploid (2n = 48) M. citrina four 45S rDNA and five 5S rDNA loci have been detected. No single chromosome of M. citrina was uniformly labelled after using genomic probes from M. arborea and M. strasseri. Instead, cross-hybridization signals in M. citrina were restricted to terminal chromosome arms and NOR regions.

Conclusions

FISH results support the close taxonomic interrelationship between M. arborea and M. strasseri. In these tetraploid species, NOR loci have experienced a diploidization event through physical loss of sequences, a cytogenetic feature so far not reported in other species of the genus. The high number of rDNA loci and GISH results support the specific status for the hexaploid M. citrina, and it is suggested that this species is not an autopolyploid derivative of M. arborea or M. strasseri. Further, molecular cytogenetic data do not suggest the hypothesis that M. arborea and M. strasseri were involved in the origin of M. citrina. FISH mapping can be used as an efficient tool to determine the genomic contribution of M. citrina in somatic hybrids with other medic species.Key words: Medicago arborea, M. citrina, M. strasseri, rRNA genes, 18S-5·8S-25-S, 5S, FISH mapping, GISH, polyploidy     

不同地域乌拉尔甘草基因组的FISH分析与染色体识别

在核型分析与染色体识别基础上,分别以番茄45S和5S rDNA为探针,对3种不同地域的乌拉尔甘草进行FISH分析.结果表明:内蒙古鄂托克前旗的乌拉尔甘草核型公式为2n=2x=16=6m+10sm (2SAT),新疆阿勒泰地区的乌拉尔甘草核型公式为2n=2x=16=4m+12sm(2SAT),内蒙古喀喇沁旗乌拉尔甘草核型公式为2n=2x=16=4m+12sm(2SAT);其第8染色体均带有随体.3种乌拉尔甘草基因组内均有1对5S rDNA和1对45S rDNA杂交位点.核型分析显示,5S rDNA杂交位点均位于第2染色体的短臂部位,45S rDNA杂交位点均位于第8染色体的次缢痕和随体部位.45S与5S rDNA在3种乌拉尔甘草中期分裂相上的位点数和分布情况高度一致,表明来自3种不同地域的乌拉尔甘草在染色体结构水平上没有较大的分化.     

Molecular cytogenetic analysis of Siberian Larix species by fluorescence in situ hybridization

Karyotypes of three Larix species (L. sibirica, L. gmelinii, and L. cajanderi) were analyzed using fluorescence in situ hybridization (FISH) with 45S and 5S ribosomal RNA gene probes and 4′,6-diamidino-2-phenylindole (DAPI) staining. Two major 45S ribosomal DNA (rDNA) loci (per haploid genome) have been observed in the intercalary regions of two metacentric chromosomes, III and IV, of L. sibirica; in addition to them, minor nucleolus organizing regions (NORs) were mapped in pericentromeric regions of chromosomes I, II, VI, and XII. Two closely related species, L. gmelinii and L. cajanderi, showed similar hybridization patterns; both species possessed an additional major locus of 45S rDNA in the distal region of the long arm of submetacentric chromosome VII that is absent in L. sibirica. Only one locus of the 5S rDNA was found in all larch species we studied; it was located in the distal region of the chromosome III short arm, which also carried the major NOR in the opposite arm. This chromosome containing major loci of the two ribosomal RNA gene families can serve as a marker of the genus Larix. The intra- and interspecific karyotype diversity in the genus Larix is discussed.     

Polymorphisms in bovine immune genes and their associations with somatic cell count and milk production in dairy cattle

Background

Chromosome rearrangements are an important part of the speciation process in many taxa. The study of chromosome evolution in bivalves is hampered by the absence of clear chromosomal banding patterns and the similarity in both chromosome size and morphology. For this reason, obtaining good chromosome markers is essential for reliable karyotypic comparisons. To begin this task, the chromosomes of the mussels Brachidontes puniceus and B. rodriguezi were studied by means of fluorochrome staining and fluorescent in situ hybridization (FISH).

Results

Brachidontes puniceus and B. rodriguezi both have 2n = 32 chromosomes but differing karyotype composition. Vertebrate-type telomeric sequences appear at both ends of every single chromosome. B. puniceus presents a single terminal major rRNA gene cluster on a chromosome pair while B. rodriguezi shows two. Both mussels present two 5S rDNA and two core histone gene clusters intercalary located on the long arms of two chromosome pairs. Double and triple-FISH experiments demonstrated that one of the 5S rDNA and one of the major rDNA clusters appear on the same chromosome pair in B. rodriguezi but not in B. puniceus. On the other hand, the second 5S rDNA cluster is located in one of the chromosome pairs also bearing one of the core histone gene clusters in the two mussel species.

Conclusion

Knowledge of the chromosomal distribution of these sequences in the two species of Brachidontes is a first step in the understanding of the role of chromosome changes on bivalve evolution.     

Cytogenetic characterization of the sole Solea senegalensis (Teleostei: Pleuronectiformes: Soleidae): Ag-NOR, (GATA) n , (TTAGGG) n and ribosomal genes by one-color and two-color FISH

A cytogenetic analysis of the sole Solea senegalensis was carried out using silver staining for the nucleolus organizer region (Ag-NOR) identification, one-color FISH for chromosomal mapping of 45S and 5S ribosomal DNAs (rDNAs), (GATA) _n , and (TTAGGG) _n , and two-color FISH for co-localization of both rDNAs. The Ag-NORs and the 45S rDNA were mapped to a medium-sized submetacentric chromosomal pair. Hybridization with the 5S rDNA showed a major signal on the short arm of a medium-sized submetacentric chromosome pair and a minor signal on a centromeric site of a small acrocentric chromosome pair. Differences in the Ag-NOR and 45S and 5S rDNAs FISH signal sizes were observed between homologous chromosomes and among individuals. A two-color FISH co-localized 45S and 5S rDNAs to a medium-sized submetacentric chromosomal pair. The hybridization with the telomeric (TTAGGG) _n repeat displayed small signals at all chromosomal telomeres. Finally, the (GATA) _n probe produced dispersed and small hybridization signals on all chromosome spreads, showing its ubiquitous existence in the genome. These results were compared with those from other Pleuronectiformes and discussed in terms of karyotype evolution.