Physical mapping of the 5S ribosomal RNA genes in Arabidopsis thaliana by multi-color fluorescence in situ hybridization with cosmid clones

The 5S ribosomal RNA genes were mapped to mitotic chromosomes of Arabidopsis thaliana by fluorescence in situ hybridization (FISH). In the ecotype Landsberg erecta, hybridization signals appeared on three pairs of chromosomes, two of which were metacentric and the other acrocentric. Hybridization signals on one pair of metacentric chromosomes were much stronger than those on the acrocentric and the other pair of metacentric chromosomes, probably reflecting the number of copies of the genes on the chromosomes. Other ecotypes, Columbia and Wassilewskija, had similar chromosomal distribution of the genes, but the hybridization signals on one pair of metacentric chromosomes were very weak, and detectable only in chromosomes prepared from young flower buds. The chromosomes and arms carrying the 5S rDNA were identified by multi-color FISH with cosmid clones and a centromeric 180 bp repeat as co-probes. The metacentric chromosome 5 and its L arm carries the largest cluster of the genes, and the short arm of acrocentric chromosome 4 carries a small cluster in all three ecotypes. Chromosome 3 had another small cluster of 5S rRNA genes on its L arm. Chromosomes 1 and 2 had no 5S rDNA cluster, but they are morphologically distinguishable; chromosome 1 is metacentric and 2 acrocentric. Using the 5S rDNA as a probe, therefore, all chromosomes of A. thaliana could be identified by FISH. Chromosome 1 is large and metacentric; chromosome 2 is acrocentric carrying 18S-5.8S-25S rDNA clusters on its short arm; chromosome 3 is metacentric carrying a small cluster of 5S rDNA genes on its L arm; chromosome 4 is acrocentric carrying both 18S-5.8S-25S and 5S rDNAs on its short (L) arm; and chromosome 5 is metacentric carrying a large cluster of 5S rDNA on its L arm.     

Molecular cytogenetics of <Emphasis Type="Italic">Oligosarcus hepsetus</Emphasis> (Teleostei,Characiformes) from two Brazilian locations

Karyotype and cytogenetic markers of Oligosarcus hepsetus from two Brazilian locations in the Paraíba do Sul River Basin (Brazil) were investigated using differential staining techniques (C-banding, silver (Ag)- and chromomycin A₃ (CMA₃)-staining) and fluorescent in situ hybridization (FISH) using 18 S rDNA and 5 S rDNA probes. The diploid chromosome number was invariably 2n = 50 with 3 pairs of metacentric, 5 pairs of submetacentric, 8 pairs of subtelocentric and 9 pairs of acrocentric chromosomes. No heteromorphic sex chromosomes were observed. The nucleolar organizer regions (NORs) were detected in the short arms of the largest acrocentric pair using Ag-, CMA₃- stainings and FISH with 18 S rDNA probe, the latter showing also positive labeling in the short arms of a small acrocentric pair, not visualized by the former methods. FISH with 5 S rDNA probe showed positive labeling in the two chromosome pairs. While the CMA₃-staining exhibited GC-rich heterochromatin segments in two pairs of chromosomes, including those coincided with Ag-NORs, the DAPI staining did not reveal any signal, indicating the absence of AT-rich heterochromatin. FISH with an As-51 satellite DNA probe derived from the closely related Astyanax scabripinnis did not reveal any positive signal, demonstrating the absence of this class of DNA in the genome of the specimens under study.     

Chromosome number and ploidy-level in a Dutch population ofCrenobia alpina Dana (Planaria)

A Dutch population ofCrenobia alpina was found to have a chromosome number of 2n=42. One pair of large chromosomes is acrocentric and probably shows a secondary constriction; one small pair is acrocentric as well. All other chromosomes are metacentric or sub-metacentric; it has not been possible to discriminate with certainty between them. According to the hypothesis ofDahm (1958) populations with, 2n=42 are autohexaploid, the basic genome consisting of seven chromosomes. On basis of the karyotype it was concluded that the Dutch population described here, is not autohexaploid but functionally diploid, perhaps allohexaploid.     

Characterization of a complex restriction-modification system detected in<Emphasis Type="Italic">Staphylococcus aureus</Emphasis> and<Emphasis Type="Italic">Streptococcus agalactiae</Emphasis> strains isolated from infections of domestic animals

Characterization of classic type II restriction-modification systems (RMS) (restriction endonucleases and modification methyltransferases) was carried out in isolates ofStaphylococcus aureus andStreptococcus agalactiae obtained from clinical material. Among the 100 isolates ofS. aureus two different RMS type II were detected. The first was expressed in isolates 32 and 33 (Sau32 I andSau33 I); the targeting sequence was determined as 5′-GGN CC-3′ (Sau96 I isoschizomer). The second was found in isolates no. 90, 93, 96*, and 98 (Sau90 I,Sau93 I,Sau96* I,Sau98 I) and enzymes recognized sequence 5′-CTY RAG-3′ (SmlI isoschizomer). Analysis of 40 isolates ofS. agalactiae revealed only one RMS; it was detected in two isolates (no. 16 and 23;Sag16 I andSag23 I). Restriction endonuclease expressed by these isolates cleaved DNA in sequence 5′-CTG CA/G-3′ (PstI isoschizomer). In RMS-positiveS. aureus andS. agalactiae isolates plasmid DNA capable of replication inEscherichia coli andBacillus subtilis was also detected and isolated. This research was supported by VEGA grant of theSlovak Academy of Sciences no. 2/2059/22 and grant no. 2003 SP27/0208E 02/028/0E02 of theMinistry of Agriculture of the Slovak Republic.     

The karyotype and 5S rRNA genes from Spanish individuals of the bat species <Emphasis Type="Italic">Rhinolophus</Emphasis><Emphasis Type="Italic">hipposideros</Emphasis> (Rhinolophidae; Chiroptera)

The karyotype of individuals of the species Rhinolophus hipposideros from Spain present a chromosome number of 2n = 54 (NFa = 62). The described karyotype for these specimens is very similar to another previously described in individual from Bulgaria. However, the presence of one additional pair of autosomal acrocentric chromosomes in the Bulgarian karyotype and the differences in X chromosome morphology indicated that we have described a new karyotype variant in this species. In addition, we have analyzed several clones of 1.4 and 1 kb of a PstI repeated DNA sequence from the genome of R. hipposideros. The repeated sequence included a region with high identity with the 5S rDNA genes and flanking regions, with no homology with GenBank sequences. Search for polymerase III regulatory elements demonstrated the presence of type I promoter elements (A-box, Intermediate Element and C-box) in the 5S rDNA region. In addition, upstream regulatory elements, as a D-box and Sp1 binding sequences, were present in flanking regions. All data indicated that the cloned repeated sequences are the functional rDNA genes from this species. Finally, FISH demonstrated the presence of rDNA in nine chromosome pairs, which is surprising as most mammals have only one carrier chromosome pair.     

Chromosomal diversification of reef fishes from genus <Emphasis Type="Italic">Centropyge</Emphasis>(Perciformes,Pomacanthidae)

The genus Centropyge is remarkable for species richness, composing a highly specialized fish group amongst members from family Pomacanthidae. However, cytogenetical reports are nearly absent in these animals. New data are provided from karyotypical studies carried out on Centropyge aurantonotus from the Brazilian coast of the Atlantic Ocean and C. ferrugatus from the Philippines Sea of the Indo-Pacific Ocean. Both species present 2n=48 but karyotypes are differentiated by fundamental number. C. aurantonotus has a great number of biarmed chromosomes (4 m + 14 sm+16 st+4 a), while C. ferrugatus presents only acrocentric chromosomes. Single nucleolar organizer regions (NORs) are located at interstitial position of an acrocentric pair in C. ferrugatus and on short arms of a subtelocentric pair in C. aurantonotus, as confirmed by fluorescent in situ hybridization (FISH) with 18S rDNA probes. Heterochromatin is distributed over NOR and centromeric regions in both species, but additional GC-rich heterochromatic blocks on short arms of up to eight chromosomal pairs can be detected in C. aurantonotus. 5S rDNA segments were located interstitially on two chromosomal pairs in C. ferrugatus and on nine pairs in C. aurantonotus, mostly equivalent to heterochromatic blocks on short arms of biarmed chromosomes. C. ferrugatus can be considered a species in which basal chromosomal features proposed for modern Teleosteans were conserved. The derived karyotype pattern of C. aurantonotus seems to be determined by pericentric inversions and heterochromatin addition which probably determined the notorious dispersion of 5S rRNA (pseudo)genes. It is demonstrated that, even within a group generally characterized by cytogenetical homogeneity as the family Pomacanthidae, diversified karyotypes can be found.     

Karyological study of four Japanese Myotis bats (Chiroptera,Mammalia)

Karyological investigations of four Japanese Myotis species were made based on Gand C-banding pattern analysis. It was revealed that the four species, M. nattereri, M. hosonoi, M. frater kaguyae and M. macrodactylus have all 2n=44 and their karyotypes are, excepting one chromosome pair, identical each other. The only difference in their karyotypes was found on the morphology of the chromosome no. 5. A minute acrocentric (A) was observed in M. nattereri, and a polymorphic (A) and an (M^h) which is a minute metacentric with totally heterochromatic arm was found in M. hosonoi. In M. f. kaguyae, pair no. 5 was a small submetacentric with a totally heterochromatic long arm (SM^h). Polymorphic (SM^h) and (M) which is a small metacentric derived from (SM^h) by a pericentric inversion was seen in M. macrodactylus. Such morphological differentiations of no. 5 were interpreted by assuming an increase of constitutive heterochromatin and also an inversion. The evolutionary pathway in the genus Myotis is assumed to be as follows: (A)(M^h)(SM^h)(M). This assumption was supported by the geographical evidence that the species with the (A) type no. 5 pair is widely distributed in the whole world but the others are restricted to Asia (M^h type) or only to Japan (SM^h and M types).     

Chromosomal description of the rodent generaOecomys andNectomys from Brazil

We analysed karyotypes of five taxa of the rodent generaOecomys andNectomys, trapped in 14 localities in an area ranging from 8° to 29°S on Brazilian territory.Oecomys cf.concolor, collected in the Amazon and in two localities of the Cerrado biome, showed a 2n=60 karyotype constituted by a pair of large subtelocentric chromosomes, a small metacentric pair and 27 acrocentric pairs. The X chromosome was a large submetacentric and a subtelo-submetacentric, the morphology of the latter showing variable C-banding patterns. In all three localities the Y chromosome was a medium size heterochromatic acrocentric. Two individuals from the Cerrado had a heterochromatic acrocentric B-chromosome.Oecomys cf.bicolor presented two cytotypes, 2n=80 in the specimens from the Cerrado biome and 2n=82 in individuals trapped in the Amazon. The 2n=80 cytotype 1 showed a large subtelocentric, 22 biarmed pairs (medium to small) and 16 acrocentric autosomal pairs. The karyotype of the 2n=82 cytotype 2 is constituted by 15 biarmed chromosomes (median to small) and 25 acrocentric pairs with heterochromatic blocks at pericentromeric regions. The sexual pairs were the same (large submetacentric X and median acrocentric Y) in both cytotypes. InO. cf.concolor and in both cytotypes ofO. cf.bicolor the nucleolar organizer regions were observed in 1-3 pairs, located in the short arms.Nectomys genus presented two cytotypes, 2n=52–55 (N. rattus, with 0–3 biarmed heterochromatic accessory chromosomes) and 2n=56–59 (N. squamipes, bearing 0–3 biarmed, heterochromatic, B-chromosomes). These 2 cytotypes occupy disjunct regions of South America, with overlapping areas in the Brazilian states of Pernambuco, Bahia, and Mato Grosso do Sul.     

Sedum surculosum andS. jaccardianum (Crassulaceae) share a unique 70 bp deletion in the chloroplast DNA trnL (UAA)-trnF (GAA) intergenic spacer

TrnL (UAA)-trnF (GAA) chloroplast DNA spacer sequences of three species ofMonanthes, Sedum surculosum (=Monanthes atlanticum) andS. jaccardianum were compared.S. surculosum, the systematic position of which has been disputed ever since its discovery, shares a phylogenetically highly significant 70 bp deletion withS. jaccardianum. In addition to this large deletion the two Moroccan species ofS. ser.Monanthoidea differ in three more indels as well as in four nucleotide substitutions from the species ofMonanthes. These data render strong support for the monophyly ofS. ser.Monanthoidea andMonanthes. Spacer length in seven species and one subspecies ofMonanthes is relatively uniform.     

Karyotype of the Snow Sculpin <Emphasis Type="Italic">Myoxocephalus brandti</Emphasis> Steindachner (Cottidae) from Peter the Great Bay,Sea of Japan

The karyotype of the snow sculpin Myoxocephalus brandti, 2n = 44, NF = 46, from Peter the Great Bay was studied. Two-armed chromosomes were presented by one pair of metacentric chromosomes of medium size; one-armed chromosomes included two pairs of large subtelocentric chromosomes and a pair of large acrocentric chromosomes. Ag-NOR-staining in the telomere vicinity revealed nucleolus-organizing regions in one metacentric chromosome and in one medium size acrocentric chromosome in one of the fishes, in two homological small acrocentric chromosomes in three fishes, and in one acrocentric chromosome of average size in six fishes. No difference between the male and female karyotypes and any type of variability was revealed. The karyotypes of the snow sculpin M. brandti and the frog sculpin M. stelleri were compared. Their distinctions and similarities were displayed.Original Russian Text Copyright ¢ 2005 by Biologiya Morya, Ryazanova.     

Karyological studies on Indian spiders

The diploid set of chromosomes inPalystes whiteae Pocock (Eusparassidae),Selenops montigenus Simon (Selenopidae) andTarrocanus viridis Dyal (Thomisidae) is constituted by forty-three, twenty-nine and twenty-seven acrocentric elements respectively. The males of all these three species have a sex-determining mechanism of the XXXO-type. The autosomes ofP. whiteae andT. viridis are simple rods, more or less pointed at one end, while inS. montigenus they carry terminal knobs. The sex-chromosomes which are unequal, move to one pole of the spindle precociously, during anaphase I.     

Evidence for eight tandem and five centric fusions in the evolution of the karyotype ofAethomys namaquensis A. Smith (Rodentia: Muridae)

G- and C-banded chromosomes ofAethomys namaquensis (2n=24),A. chrysophilus (2n=44), andPraomys coucha (2n=36) are compared and contrasted with publised material on Australian Muridae and North American Sigmodontidae. Direction and types of chromosomal rearrangements are established using cladistic methodology. An acrocentric morphology for chromosomes 5, 14, 15 and 20 (numbering system fromPeromyscus) are proposed as primitive for the common ancestor of the Muridae and Sigmodontidae rodent lineages. Reduced diploid number ofAethomys namaquensis is derived by eight tandem and five centric fusions since divergence from the common ancestor withA. chrysophilus. The two species ofAethomys share one derived metacentric chromosome that distinguishes them fromPraomys. Praomys has unique chromosomes which can be derived from the proposed primitive condition by five centric fusions and five pericentric inversions. It is concluded that karyotypic orthoselection for tandem and centric fusions is best explained by cellular or biochemical mechanisms rather than variation in population characteristics.     

Karyological studies on the Indian spiders VII. Mitosis and meiosis in two species belonging to the family gnaphosidae

Gnaphosa kailana Tikader andScotophaeus blackwallii (Thorell) are characterized by twenty-two (20+X₁X₂) and twenty-four (22+X₁X₂) acrocentric chromosomes in their diploid set respectively. In case ofG. kailana the sex-chromosomes are slightly unequal while inS. blackwallii the size difference is well marked between the two. The sex-chromosomes in both species form an accessory plate at the equator of the spindle during metaphase-I and show a precocious anaphase-I movement.     

Taxonomic reappraisal on<Emphasis Type="Italic">Suaeda australis</Emphasis> (Chenopodiaceae) in korea based on the morphological and molecular characteristics

We used morphological and molecular characteristics to perform a taxonomic reappraisal ofSuaeda australis (Brown) Moquin-Tandon from Korea. Populations of this species are dispersed at the bottoms of sand zones, and usually exhibit a depressed habit. Except for their total heights and leaf lengths, the morphology of these plants does not differ from that ofS. maritima. Molecular traits were examined based on ITS and psbB-psbH spacer region sequences. The former region included ITS-1, 5.8S, and ITS-2, which were 629 nucleotide bases long. Pair-wise distances (p-distance) among KoreanSuaeda species ranged from 1.12 to 17.84. The psbB-psbH spacer region sequences were 618 nucleotide bases long, and were conserved in the alignment of KoreanSuaeda species. In our ML and MrBayesian analysis of ITS sequences aligned with other sequences from GenBank, the plants of KoreanSuaeda made three clades: 1)S. japonica;S. australis, andS. maritima;2)S. malacosperma; and 3) S.glauca. However, the psbB-psbH region sequences could not be resolved amongS. japonica, S. maritima, andS. australis from Korea. Molecular and vegetative characteristics indicated that the plants now classified asS. australis from Korea should instead be named as S.maritima (L.) Dumont.     

Karyotype studies in seven species of cyprinidae

The present report embodies karyotype studies of seven Indian cyprinids. Of these, the karyotypes of five species possess chromosomes all of which are acrocentric. These areLabeo calbasu (Ham.) 2n=54,Puntius sophore (Ham.),P. conchonius Ham.,P. stigma (Ham.) 2n=50 andChela bacaila (Ham.) 2n=52. The karyotype ofCrossocheilus latius punjabensis Mukerji has 48 chromosomes, 12 of which are metacentric (2n=12m+36r's). InAmblypharyngodon mola (Ham.), 52 chromosomes form the diploid set which include 8 metacentrics. In none of the seven species morphologically distinguishable sex chromosomes were found.Literature on cyprinid karyotypes has been reviewed in order to understand in how far the cytological characters are in agreement with the classification based on morphological studies. The validity ofMatthey's notion of nombre fondamental to establish kinship between cytologically known cyprinid species has also been discussed.     

Preliminary analysis of numerical taxonomy of the genusSaguinus based on dental measurements

Although several investigations have been made from different viewpoints, the classification or interspecific relationships ofSaguinus still remain uncertain. In the present study, we applied multivariate analysis methods to dental measurements of part ofSaguinus populations of sufficient sample size and obtained the following conclusions.Saguinus can be classified into two main groups: one consists ofS. oedipus andS. leucopus, and the other ofS. fuscicollis, S. nigricollis, S. labiatus, andS. mystax. Concerning the former group, the two subspecies ofS. oedipus, S. o. oedipus, andS. o. geoffroyi, show a close affinity with each other and also a close relationship toS. leucopus, while the latter group consists of two subgroups of species, one includingS. fuscicollis andS. nigricollis, and the other includingS. labiatus andS. mystax. The biological distance betweenS. oedipus oedipus andS. o. geoffroyi is slightly larger than that between the pairs ofS. fuscicollis andS. nigricollis and ofS. labiatus andS. mystax. Factor analysis revealed significant factors which could explain the differences among the seven maleSaguinus populations. Taking all the results into account, it seems necessary to reconsider the phylogenetic relationships within the genusSaguinus.     

Resistance Transfer Factors in sensitive strains ofS. panama

Two natural strains ofS.panama with an incomplete Resistance Factor (R factor) are described:S.panama I carries a Resistance Transfer Factor (RTF) exerting restriction on phageS.panama 47, but no resistance determinants; andS.panama 219 has a tetracycline-resistance determinant but no RTF.A number of drug-sensitive strains ofS.panama belonging to various phage types, were found to carry a factor which is able to mobilise the tetracycline-resistance determinant ofS.panama 219 and also of one strain ofE.coli and to transfer them toS.panama andE.coli K 12. This factor is not identical with the F factor, it is not a bacteriocinogenic factor and can therefore be considered as an RTF. These RTFs exert no restriction on (spp) and phage 47, and some of them are fi whilst others are fi.     

Distribution of similar self-incompatibility (<Emphasis Type="Italic">S</Emphasis>) haplotypes in different genera,<Emphasis Type="Italic"> Raphanus</Emphasis> and<Emphasis Type="Italic"> Brassica</Emphasis>

The nucleotide sequences of ten SP11 and nine SRK alleles in Raphanus sativus were determined, and deduced amino acid sequences were compared with those of Brassica SP11 and SRK. The amino acid sequence identity of class-I SP11s in R. sativus was about 30% on average, the highest being 52.2%, while that of the S domain of class-I SRK was 77.0% on average and ranged from 70.8% to 83.9%. These values were comparable to those of SP11 and SRK in Brassica oleracea and B. rapa. SP11 of R. sativus S-21 was found to be highly similar to SP11 of B. rapa S-9 (89.5% amino acid identity), and SRK of R. sativus S-21 was similar to SRK of B. rapa S-9 (91.0%). SP11 and SRK of R. sativus S-19 were also similar to SP11 and SRK of B. oleracea S-20, respectively. These similarities of both SP11 and SRK alleles between R. sativus and Brassica suggest that these S haplotype pairs originated from the same ancestral S haplotypes.     

Gene mapping of 28S and 5S rDNA sites in the spined loach Cobitis taenia (Pisces, Cobitidae) from a diploid population and a diploid–tetraploid population

We compare the chromosomal 28S and 5S rDNA patterns of the spined loach C. taenia (2n = 48) from an exclusively diploid population and from a diploid–polyploid population using 28S and 5S rDNA probe preparation and labelling, and fluorescence in situ hybridization (FISH). The 5S rDNA was located in two to three chromosome pairs, and separated from the 28S loci for the males and one female (F1) from the diploid population. Loaches from a diploid–polyploid population, and one female (F2) from the diploid population were characterized by at least one chromosome pair with 5S and 28S overlapping signals. The fishes differed mainly in their number of 28S rDNA loci, located on 3–6 chromosomes. All individuals from both populations were characterized by one acrocentric chromosome bearing a 28S rDNA signal on the telomeres of its long arm. The number of major ribosomal DNA in the karyotype of C. taenia by FISH was always higher than the number of Ag-NORs. Our data confirm the extensive polymorphism of NORs in both populations, as already has been observed in closely related Cobitis species, and less polymorphic 5S rDNA pattern. However, this preliminary result highlights the need for a wider scale study.     

Epibiota communities of the introduced and indigenous macroalgal relatives<Emphasis Type="Italic"> Sargassum muticum</Emphasis> and<Emphasis Type="Italic"> Halidrys siliquosa</Emphasis> in Limfjorden (Denmark)

Sargassum muticum (Phaeophyceae, Fucales) has recently been introduced to Limfjorden (Denmark) where its closest relative is the indigenous Halidrys siliquosa. Previous studies have demonstrated large quantitative (canopy biomass) and qualitative (canopy persistence) differences in the habitat available to epibiota within the canopies of these two macroalgae. We therefore hypothesised that these algae would support different epibiota communities and tested this by sampling the epibiota of S. muticum and H. siliquosa on seven occasions throughout 1997 by enclosing entire thalli in mesh bags. We found 53 epibiota taxa and, with only one exception, they were all recorded on both host species. Species richness and abundance of epibiota exhibited clear seasonal variation on both host species, although epibiota biomass was seasonally constant on H. siliquosa but not on S. muticum. These patterns were consistent with the different life histories of the host species. There was a weakly negative correlation between thallus size and epibiota biomass for both host species. When taking species-specific seasonal variation in thallus size into consideration, S. muticum and H. siliquosa were found to support significantly different epibiota biomasses. Multivariate analyses showed that epibiota community structure was different, although highly overlapping, between the two species, whereas there was an almost parallel temporal development in epibiota community structure. We conclude that it is unlikely that the introduction of S. muticum to Limfjorden has caused major changes in local epibiota community structure. However, the standing stock of epibiota is likely to have increased.Communicated by H.-D. Franke