Chromosomal distribution of 18S-25S rDNA in four <Emphasis Type="Italic">Lupinus</Emphasis> species visualized by fluorescence <Emphasis Type="Italic">in situ</Emphasis> hybridization

The number and position of 18S–25S rDNA sites in 4 selected Lupinus species are reported for the first time. L. atlanticus, L. subcarnosus and L. paniculatus had two rDNA loci, while L. albus exhibited only one loci. Among these 4 species, all of them exhibited one large pair of strong signals that extends from the short arm to a NOR on a chromosome satellite. L. atlanticus, L. subcarnosus, L. paniculatus had one more locus of 18–25S rDNA, but a pair of weak hybridization signals were observed in L. paniculatus when 18S–25S rDNA was used as probe. The results are discussed in terms of the evolutionary relationships among these species.     

Distribution of 5S and 18S–28S rDNA loci in a tetraploid cotton (Gossypium hirsutum L.) and its putative diploid ancestors

The most widely cultivated species of cotton,Gossypium hirsutum, is a disomic tetraploid (2n=4x=52). It has been proposed previously that extant A- and D-genome species are most closely related to the diploid progenitors of the tetraploid. We used fluorescent in situ hybridization (FISH) to determine the distribution of 5S and 18S-28S rDNA loci in the A-genome speciesG. herbaceum andG. arboreum, the D-genome speciesG. raimondii andG. thurberi, and the AD tetraploidG. hirsutum. High signal-to-noise, single-label FISH was used to enumerate rDNA loci, and simultaneous, dual-label FISH was used to determine the syntenic relationships of 5S rDNA loci relative to 18S–28S rDNA loci. These techniques provided greater sensitivity than our previous methods and permitted detection of six newG. hirsutum 18S–28S rDNA loci, bringing the total number of observed loci to 11. Differences in the intensity of the hybrizization signal at these loci allowed us to designate them as major, intermediate, or minor 18–28S loci. Using genomic painting with labeled A-genome DNA, five 18S–28S loci were localized to theG. hirsutum A-subgenome and six to the D-subgenome. Four of the 11 18S–28S rDNA loci inG. hirsutum could not be accounted for in its presumed diploid progenitors, as both A-genome species has three loci and both D-genome species had four.G. hirsutum has two 5S rDNA loci, both of which are syntenic to major 18S–28S rDNA loci. All four of the diploid genomes wer examined contained a single 5S locus. InG. herbaceum (A₁) andG. thurberi (D₁), the 5S locus is syntenic to a major 18S–28S locus, but inG. arboreum (A₂) andG. raimondii (D₅), the proposed D-genome progenitor ofG. hirsutum, the 5S loci are syntenic tominor and intermediate 18S–28S loci, respecitively. The multiplicity, variation in size and site number, and lack of additivity between the tetraploid species and its putative diploid ancestors indicate that the behavior of rDNA loci in cotton is nondogmatic, and considerably more complex and dynamic than previously envisioned. The relative variability of 18S–28S rDNA loci versus 5S rDNA loci suggests that the behavior of tandem repearts can differ widely. Edited by: R. Appels     

Comparative mapping between<Emphasis Type="Italic"> Quercus</Emphasis> and<Emphasis Type="Italic"> Castanea</Emphasis> using simple-sequence repeats (SSRs)

Simple sequence repeat (SSR) markers from Quercus and Castanea were used for comparative mapping between Quercus robur (L.) and Castanea sativa (Mill.). We tested the transferability of SSRs developed in Quercus to Castanea and vice-versa. In total, 47% (25) of the Quercus SSRs and 63% (19) of the Castanea SSRs showed a strong amplification product in the non-source species. From these 44 putative comparative anchor tags, 19 (15 from Quercus and 4 from Castanea) were integrated in two previously established genetic linkage maps for the two genera. SSR loci were sequenced to confirm the orthology of the markers. The combined information from both genetic mapping and sequence analysis were used to determine the homeology between seven linkage groups, aligned on the basis of pairs or triplets of common markers, while two additional groups were matched using a single microsatellite marker. Orthologous loci identified between Q. robur and C. sativa will be useful as anchor loci for comparative mapping studies within the Fagaceae family.Communicated by D.B. NealeThis paper is dedicated to the memory of Paulo Costa     

Cytological heterozygosity and the hybrid origin of sweet orange [Citrus sinensis (L.) Osbeck]

Citrus sinensis chromosomes, although small in size, present a remarkable differentiation of bands with the fluorochromes CMA and DAPI. These bands suggest that some heteromorphisms are fixed in this species. To investigate the extension of these heteromorphisms, ten cultivars of C. sinenesis were analysed with CMA/DAPI staining and, in some of them, the 18S–5.8S–25S rRNA and 5S rRNA genes were located by in situ hybridization. CMA/DAPI staining showed exactly the same CMA⁺/DAPI⁻ banding pattern for all cultivars. In situ hybridization revealed three 18S–5.8S–25S rRNA gene sites, two proximally located on two similar chromosomes and one terminally located on a third non-related chromosome. Two 5S rRNA gene sites were observed in this species, with one located proximal to the telomeric 18S–5.8S–25S rDNA site. Both cytological approaches revealed an invariable, heterozygotic karyotype among sweet orange cultivars. Based on these data, the putative hybrid origin of the species is discussed. Received: 9 April 1999 / Accepted: 22 June 1999     

Karyotypes of three somaclonal variants and wild plants ofAllium tuberosum by bicolor FISH

Using the fluorescence in situ hybridization (FISH) technique, we conducted karyotype analyses to identify the lost chromosomes in three somaclonal variants obtained from tissue culture of wildAllium tuberosum (2n = 4X = 32). The three lost chromosomes of the At29 variant (2n = 29) were all chromosome 2, the two for At30 (2n = 30) were chromosomes 7 and 8, and At31 was missing chromosome 2. Chromosome compositions of these variants were confirmed as being fixed lines during two years of greenhouse cultivation. The bicolor FISH technique, involving both 5S and 18S–5.8S–26S ribosomal RNA genes as probes, was used to assign chromosomal locations and to confirm whether the lost chromosomes contained any rRNA markers. The 5S rRNA gene signals in all variants as well as the wild type were detected as two sets, one on the intercalary region of the short arm of chromosome 3, the other on the intercalary region of the long arm of chromosome 6. One 18S–5.8S–26S rRNA gene site on the secondary constriction included a flanking satellite and terminal region on the short arm of chromosome 8. Signals of the 18S–5.8S–26S rRNA gene in At30 showpd in only three chromosomes, indicating that one of the lost chromosomes was chromosome 8. Overall, three marker chromosomes were established by FISH, using rRNA multigene families.     

Mapping of ribosomal DNA and (TTAGGG)n telomeric sequence by FISH in the bivalve Patinopecten yessoensis (Jay, 1857)

The chromosome set of Patinopecten yessoensis (Jay, 1857) wascharacterized using Giemsa staining, DAPI staining and fluorescencein situ hybridization (FISH) with three repetitive DNA probes[18S–28S rDNA, 5S rDNA and telomeric (TTAGGG)_n]. DAPIstaining showed that AT-rich regions were located on the centromereof almost all chromosomes and interstitial banding was not observed.FISH showed that 18S–28S rDNA spread over the short armsof two subtelocentric chromosome pairs and 5S rDNA was locatedon the long arm of one subtelocentric chromosome pair. SequentialFISH demonstrated that 18S–28S and 5S rDNA were locatedon different chromosomes. FISH also showed that the vertebratetelomeric sequence (TTAGGG)_n was located on both ends of eachchromosome and no interstitial signals were detected. Sequential18S–28S rDNA and (TTAGGG)_n FISH indicated that repeatedunits of the two multicopy families were closely associatedon the same chromosome pair. (Received 4 January 2007; accepted 1 September 2007)     

Chromosomal polymorphism of ribosomal genes in the genus Oryza

The genes encoding for 18S–5.8S–28S ribosomal RNA (rDNA) are both conserved and diversified. We used rDNA as probe in the fluorescent in situ hybridization (rDNA-FISH) to localized rDNAs on chromosomes of 15 accessions representing ten Oryza species. These included cultivated and wild species of rice, and four of them are tetraploids. Our results reveal polymorphism in the number of rDNA loci, in the number of rDNA repeats, and in their chromosomal positions among Oryza species. The numbers of rDNA loci varies from one to eight among Oryza species. The rDNA locus located at the end of the short arm of chromosome 9 is conserved among the genus Oryza. The rDNA locus at the end of the short arm of chromosome 10 was lost in some of the accessions. In this study, we report two genome specific rDNA loci in the genus Oryza. One is specific to the BB genome, which was localized at the end of the short arm of chromosome 4. Another may be specific to the CC genome, which was localized in the proximal region of the short arm of chromosome 5. A particular rDNA locus was detected as stretched chromatin with bright signals at the proximal region of the short arm of chromosome 4 in O. grandiglumis by rDNA-FISH. We suggest that chromosomal inversion and the amplification and transposition of rDNA might occur during Oryza species evolution. The possible mechanisms of cyto-evolution in tetraploid Oryza species are discussed.     

Comparative physical mapping of the 5S and 18S-25S rDNA in nine wild Hordeum species and cytotypes

Absract  The physical locations of the 5S and 18S-25S rDNA sequences were examined in nine wild Hordeum species and cytotypes by double-target in situ hybridization using digoxigenin-labelled 5S rDNA and biotin-labelled 18S-25S rDNA as probes. H. vulgare ssp. spontaneum (2n=2x=14; I-genome) had a similar composition of 5S and 18S-25S rDNA to cultivated barley (H. vulgare ssp. vulgare, I-genome), with two major 18S-25S rDNA sites and minor sites on four of the other five chromosomes; three chromosomes had 5S rDNA sites. The closely related H. bulbosum (2x; also I-genome) showed only one pair of 5S rDNA sites and one pair of 18S-25S rDNA sites on different chromosomes. Four wild diploid species, H. marinum (X-genome), H. glaucum and H. murinum (Y-genomes) and H. chilense (H-genome), differed in the number (2–3 pairs), location, and relative order of 5S and the one or two major 18S-25S rDNA sites, but no minor 18S-25S rDNA sites were observed. H. murinum 4x had three chromosome pairs carrying 5S rDNA, while the diploid had only a single pair. Two other tetraploid species, H. brachyantherum 4x and H. brevisubulatum 4x (both considered to have H-type genomes), had minor 18S-25S rDNA sites, as well as the major sites. Unusual double 5S rDNA sites – two sites on one chromosome arm separated by a short distance – were found in the American H-genome species, H. chilense and H. brachyantherum 4x. The results indicate that the species H. brachyantherum 4x and H. brevisubulatum 4x have a complex evolutionary history, probably involving the multiplication of minor rDNA sites (as in H. vulgare sensu lato), or the incorporation of both I and H types of genome. The rDNA markers are useful for an investigation of chromosome evolution and phylogeny. Received: 9 February 1998 / Accepted: 14 July 1998     

Genomics of Fagaceae

An overview of recent achievements and development of genomic resources in the Fagaceae is provided, with major emphasis on the genera Castanea and Quercus. The Fagaceae is a large plant family comprising more than 900 species belonging to 8?C10 genera. Using a wide range of molecular markers, population genetics and gene diversity surveys were the focus of many studies during the past 20?years. This work set the stage for investigations in genomics beginning in the early 1990s and facilitated the application of genetic and quantitative trait loci mapping approaches. Transferability of markers across species and comparative mapping have indicated tight macrosynteny between Quercus and Castanea. Omic technologies were more recently developed and the corresponding resources are accessible via electronic and physical repositories (expressed sequence tag sequences, single-nucleotide polymorphisms, candidate genes, cDNA clones, bacterial artificial chromosome (BAC) libraries) that have been installed in North America and Europe. BAC libraries and physical maps were also constructed in Castanea and Quercus and provide the necessary resources for full nuclear genome sequencing projects that are currently under way in Castanea mollissima (Chinese chestnut) and Quercus robur (pedunculate oak).     

Carbon allocation to defense, storage, and growth in seedlings of two temperate broad-leaved tree species

Optimal carbon allocation to growth, defense, or storage is a critical trait in determining the shade tolerance of tree species. Thus, examining interspecific differences in carbon allocation patterns is useful when evaluating niche partitioning in forest communities. We hypothesized that shade-tolerant species allocate more carbon to defense and storage and less to growth compared to shade-intolerant species. In gaps and forest understory, we measured relative growth rates (RGR), carbon-based defensive compounds (condensed tannin, total phenolics), and storage compounds (total non-structural carbohydrate; TNC) in seedlings of two tree species differing in shade tolerance. RGR was greater in the shade-intolerant species, Castanea crenata, than in the shade-tolerant species, Quercus mongolica var. grosseserrata, in gaps, but did not differ between the species in the forest understory. In contrast, concentrations of condensed tannin and total phenolics were greater in Quercus than in Castanea at both sites. TNC pool sizes did not differ between the species. Condensed tannin concentrations increased with increasing growth rate of structural biomass (GRstr) in Quercus but not in Castanea. TNC pool sizes increased with increasing GRstr in both species, but the rate of increase did not differ between the species. Accordingly, the amount of condensed tannin against TNC pool sizes was usually higher in Quercus than in Castanea. Hence, Quercus preferentially invested more carbon in defense than in storage. Such a large allocation of carbon to defense would be advantageous for a shade-tolerant species, allowing Quercus to persist in the forest understory where damage from herbivores and pathogens is costly. In contrast, the shade-intolerant Castanea preferentially invested more carbon in growth rather than defense (and similar amounts in storage as Quercus), ensuring establishment success in gaps, where severe competition occurs for light among neighboring plants. These contrasting carbon allocation patterns are closely associated with strategies for persistence in these species’ respective habitats.     

Karyotype analysis and physical mapping of 45S rDNA in eight species of Sophora, Robinia , and Amorpha

The karyotype analysis and physical locations of 45S rDNA were carried out by means of fluorescence in situ hybridization in three species, and two forms of Sophora, two species of Robina, and one species of Amorpha. S. japonica L., S. japonica L. f. oligophylla Franch., S. japonica L. f. pendula Loud., and S. xanthantha C. Y. Ma. are all tetraploids with 2n = 28. There were four 45S rDNA sites in pericentromeric regions of two pairs of chromosomes in each of them. S. rubriflora Tsoong. is a triploid with 2n = 21, and three sites were located in each satellite of group 5 chromosomes. In R. pseudoacacia L. (2n = 2x = 22), we examined four intensive signals in telomeric regions of two pairs of satellite chromosomes. In R. hispida L. (2n = 2x = 30), there were four other signals in centromeric regions besides those like in R. pseudoacacia. Amorpha fruticosa L. has most chromosomes (2n = 40) among the eight materials, however, there were only six 45S rDNA loci and they laid in centromeric regions, and satellites of three pairs of chromosomes. 45S rDNA is a valuable chromosomal landmark in karyotype analysis. The distribution and genomic organization of rDNA in the three genera were also discussed. __________ Translated from Acta Botanica Yunnanica, 2005, 27(3): 261–268 [译自: 云南植物研究, 2005, 27(3): 261–268]     

Localization of 5S and 25S rRNA genes on Somatic and meiotic chromosomes in <Emphasis Type="Italic">Capsicum</Emphasis> species of chili pepper

The loci of the 5S and 45S rRNA genes were localized on chromosomes in five species of Capsicum, namely, an-nuum, chacoense, frutescens, baccatum, and chinense by FISH. The 5S rDNA was localized to the distal region of one chromosome in all species observed. The number of 45S rDNA loci varied among species; one in annuum, two in chacoense, frutescens, and chinense, and four in baccatum, with the exceptions that ‘CM334’ of annuum had three loci and ‘tabasco’ of frutescens had one locus. ‘CM334’-derived BAC clones, 384B09 and 365P05, were screened with 5S rDNA as a probe, and BACs 278M03 and 262A23 were screened with 25S rDNA as a probe. Both ends of these BAC clones were sequenced. FISH with these BAC probes on pachytenes from ‘CM334’ plant showed one 5S rDNA locus and three 45S rDNA loci, consistent with the patterns on the somatic chromosomes. The 5S rDNA probe was also applied on extended DNA fibers to reveal that its coverage measured as long as 0.439 Mb in the pepper genome. FISH techniques applied on somatic and meiotic chromosomes and fibers have been established for chili to provide valuable information about the copy number variation of 45S rDNA and the actual physical size of the 5S rDNA in chili.     

Identification of Two Species of Yeast-like Symbiotes in the Brown Planthopper, <Emphasis Type="Italic">Nilaparvata lugens</Emphasis>

To determine the species of the yeast-like symbionts (YLS) in the brown planthoppers (BPH), Nilaparvata lugens, YLS were first isolated and purified by ultracentrifugation from the fat bodies of BPH, and then 18S rDNA and internal transcribed spacer (ITS)–5.8S rDNA sequences of YLS were amplified with the different general primers for fungi. The results showed that the two different 18S and ITS–5.8S rDNA sequences of YLS were obtained. One 2291-bp DNA sequence, which contained 18S and ITS–5.8S rDNA, showed the high similarity to Cryptococcus and was named Cryp-Like symbiotes. Another 1248-bp DNA sequence, which contained a part of 18S and ITS–5.8S rDNA, showed the high similarity to Pichia guilliermondii and was named Pichia-Like symbiotes. It was further proved that Cryp- and Pichia-Like symbiotes existed in BPH through nested PCR with specific primers for two symbiotes and in situ hybridization analysis using digoxigenin-labeled probes. Our results showed that BPH harbored more than one species of eukaryotic YLS, which suggested that diversity of fungal endosymbiotes may be occurred in planthoppers, just like bacterial endosymbiotes.     

Aerobiology of Fagaceae pollen in Trieste (NE Italy)

The aerobiological behaviour of Fagaceae in Trieste and the correlations with the meteorological parameters were examined. Airborne pollen grains of Castanea, Fagus and Quercus were collected from 1990 to 2003 using a Hirst type spore trap. The main pollen season (MPS) takes place in April and May for Quercus and Fagus, in June and July for Castanea. The highest values occur in year 1993 for Quercus, in 1998 for Castanea and in 1992 for Fagus. The Fagaceae content of the air is mainly due to Quercus and Castanea pollen, Fagus usually having a scarce pollen shedding in Trieste. The highest counts of Fagaceae pollen grains are found from late April to mid May and are mainly due to the pollen shedding of oaks. The cumulative counts vary over the years, with a mean value of 2.719 pollen grains, a lowest total of 1.341 in 2002 and a highest total of 4.704 in 1993. No positive nor negative long-term trends in pollen shedding are found. No cyclic variations were observed. Spearman’s correlation was used to establish the relationship between the daily pollen counts and the daily meteorological data. Daily pollen concentrations present sometimes positive correlation with temperature, negative with rainfall and wind speed, and no correlation with humidity. Fagus and Quercus start dates result positively correlated between themselves. Significant correlations are found between the start of MPS and the mean and maximum temperature in March for Fagus and Quercus, and May for Castanea.     

Chromosomal Characteristics of rDNA in European Grayling <Emphasis Type="Italic">Thymallus thymallus</Emphasis> (Salmonidae)

The chromosomal characteristics, locations and variations of two classes of ribosomal DNA (5S and 18S) were studied in European grayling karyotype (Thymallus thymallus, Salmonidae). Major rDNA sites as revealed by sequential CMA₃/Ag staining and confirmed by in situ hybridization with a 18S rDNA probe were situated in two loci and were found to be polymorphic in size and displaying several distinct forms. The 5S rDNA was located by PRINS on three pairs of subtelocentric chromosomes, additional minor signal was present at the centromere of one metacentric element. 5S sites were not associated with NORs. The dosage compensation mechanism was proposed as an explanation of high frequency of lethal rDNA-deleted forms of the NOR-bearing chromosomes. Double variable pattern in the number and location of NORs supported the bi-directional evolution of salmonid rDNA loci.     

Heterochromatin and rDNA patterns in Solanum species of the Morelloid and Dulcamaroid clades (Solanaceae)

The heterochromatin distribution and the position of 18-5.8-26S, and 5S rDNA loci were determined in 13 species of Solanum of the Morelloid and Dulcamaroid clades. The CMA/DAPI staining and FISH were employed. Two types of constitutive heterochromatin were determined: CMA⁺/DAPI^? associated to NOR and CMA⁺/DAPI^? distributed as terminal bands. In the Morelloid clade, CMA⁺/DAPI^? bands were found in five species while in the Dulcamaroid clade, only S. angustifidum presented this feature. In the Morelloid clade, two to four 18-5.8-26S rDNA loci occupied terminal positions and two rDNA 5S loci were found with variable positions (terminal, intercalary, and centromeric). In the Dulcamaroid clade, two terminal 18-5.8-26S rDNA loci were detected with the exception of S. salicifolium which possessed four such loci and two to four 5S rDNA loci. Solanum crispum is the only species possessing the 5S in synteny with 18-5.8-26S rDNA loci. Karyotype features chromosome banding pattern as well as the location of ribosomal genes which varied among the species, reflecting the chromosome differentiation and evolutionary divergence. The findings obtained contributed to the development of tools that can be used for establishing chromosomic homeologies among species and hence to clarify their taxonomic relationships.     

Cytological characterization of heterochromatin and rDNA inPinus radiata andP. taeda

Fluorochrome C-banding ofPinus radiata andP. taeda metaphase chromosomes showed many pericentromeric DAPI bands and interstitial CMA bands inP. radiata, and centromeric and interstitial CMA bands inP. taeda. Giemsa C-band patterns differed between the species with centromeric bands inP. radiata but no consistent bands inP. taeda. A karyotype ofP. radiata was developed based on banding patterns that distinguished all but two of the 12 pairs of chromosomes. In situ hybridization (ISH) using probes for high-copy ribosomal DNA (rDNA) showed 10 pairs of 18S–25S sites and two pairs of 5S sites in both species. Most of the sites were interstitial or centromeric.     

ITS sequences from nuclear rDNA suggest unexpected phylogenetic relationships between Euro-Mediterranean, East Asiatic and North American taxa ofQuercus (Fagaceae)

Nucleotide sequences from the internal transcribed spacer (ITS) regions of the 18S–26S nuclear ribosomal DNA have been studied from ten species ofQuercus (representing four subgenera),Castanea sativa andFagus sylvatica, as a preliminary molecular contribution to the still poorly understood systematics and evolution ofFagaceae. The resulting matrix has been used to calculate pair-wise sequence divergence indices and to construct a maximum parsimony tree forQuercus coding indels as a fifth state. Divergence is greater forQuercus vs.Fagus than forQuercus vs.Castanea. The tree for theQuercus taxa studied reveals two clearly divergent clades. In clade I the evergreen W MediterraneanQ. suber appears in a basal position as sister to more distal deciduous taxa, i.e. the E MediterraneanQ. macrolepis and the E AsiaticQ. acutissima (all formerly united as different sections under the apparently polyphyletic subg.Cerris), andQ. rubra (a representative of the N American subg.Erythrobalanus), forming a pair withQ. acutissima. In clade II the evergreen southeastern N AmericanQ. virginiana is basal and sister to the remaining three branches, i.e. a pair of evergreen Mediterranean taxa withQ. ilex andQ. coccifera (subg.Sclerophyllodrys), the deciduous but otherwise plesiomorphic SE European/SW AsiaticQ. cerris (type species of subg.Cerris), and the related but more apomorphic European pairQ. petraea andQ. robur (subg.Quercus). These results partly conflict with current taxonomic classification but are supported by some anatomical and morphological characters. They document polyphyletic lines from evergreen to deciduous taxa and suggest Tertiary transcontinental connections within the genus.     

The effects of a fogging system on the physiological status and rooting capacity of leafy cuttings of woody species

The early responses of leafy stem cuttings of Prunus and Castanea species with differing rooting abilities were assessed in a fog system using fluorescence measurements. Different types of cuttings of each species were used: cherry Prunus (‘GiSelA 5’, Prunus cerasus × Prunus canescens—148/2) and chestnut Castanea (‘Marsol’ and ‘Maraval’, Castanea crenata × Castanea sativa). The physiological status of cuttings in the early initiation phase was compared to the rooting results. For all cuttings, fluorescence measurements revealed a close-to-optimum photochemical efficiency, indicating that physiological stress (severance, water, etc.) was minimal. In cherry, the potential photochemical efficiency (Fv/Fm) differed slightly between terminal and basal cherry cuttings, being lower in the basal ones at the time of severance. Later in the propagation process, the differences were smaller. The photochemical efficiency did not differ between two ‘difficult-to-root’ Castanea clones, nor was it dependent on the length of the cuttings. The high rooting capacity of long Castanea cuttings (50 cm) indicated that physiological stress could be minimized under a fogging system. An erratum to this article can be found at     

Physical mapping of 18S-5.8S-26S and 5S ribosomal RNA gene families in three important vetches (Vicia species) and their allied taxa constituting three species complexes

The most-important vetch species, Vicia narbonensis (narbon vetch, section Faba), Vicia villosa (hairy vetch, section Cracca) and Vicia sativa (common vetch, section Vicia) and their close relatives (often difficult to circumscribe into distinct taxa) constitute respectively, Narbonensis, Villosa and Sativa species complexes in the genus Vicia. The distribution of the 18S-5.8S-26S (18S-26S) and 5S ribosomal RNA (rRNA) gene families on the chromosomes of 19 (2n=2x=10,12,14) of the 24 species and subspecies belonging to the three species complexes, and Vicia bithynica (2n=12, section Faba) and Vicia hybrida (2n=12, section Hypechusa) was studied by fluorescence in situ hybridization (FISH) with pTa 71 (18S-26S rDNA) and pTa 794 (5S rDNA) DNA clones. Computer – aided chromosome analysis was performed on the basis of chromosome length, the arm-length ratio and the position of the hybridization signals. The positions of the four (2+2) signals of the two rRNA gene families were similar between each of the three, as well as two subspecies of V. narbonensis and Vicia johannis, respectively. Two major 18S-26S rDNA loci were found in the nucleolus organiser regions (NORs) of each of the species except V. hybrida, where it was present in two out of four SAT chromosomes. In addition to major NORs, two minor loci have been physically mapped at the centromeric regions of chromosomes of group 1 in Vicia amphicarpa, Vicia macrocarpa and V. sativa, and two NORs of group 5 in V. hybrida, and on the long arms of group 4 in V. bithynica. Two or four 5S rDNA loci, observed in the short arms of groups 2–4 and 5, and 18S-26S rDNA loci were located in different chromosomes of all the species within the Narbonensis and Villosa species complexes, and Vicia angustifolia of the Sativa species complex. In the remaining six species of the Sativa species complex, and V. bithynica and V. hybrida, the two or four 5S rDNA sites were present in chromosomes which harbor 18S-26S rRNA genes. The tandemly repeated 5S rDNA sites, located at the proximal part of the long arm of groups 3–5, were diagnostic for V. angustifolia, Vicia cordata, Vicia incisa, V. macrocarpa, Vicia nigra and V. sativa of the Sativa species complex. In V. amphicarpa of the same complex, the tandem repeats were located at the distal part of the long arms of group 3. Variability in the number, size and location of two ribosomal DNA probes could generally distinguish species within the Narbonensis and Sativa species complex, V. bithynica and V. hybrida. With respect to the four species of the Villosa species complex the karyotypes could not be identified individually on the basis of the distribution of two ribosomal gene families in three out of seven pairs of chromosomes. Received: 18 October 2000 / Accepted: 20 March 2001