Cytology of Vicia species. X. Karyotype evolution and phylogenetic implication in Vicia species of the sections Atossa, Microcarinae, Wiggersia and Vicia

Automated karyotype analyses and sequence of rDNA spacers have been analysed for the species belonging to sections Atossa, Microcarinae, Wiggersia and Vicia. Karyomorphological parameters, based on Rec, Syi and TF% indices, have been determined and evidenced that, in term of symmetry, the karyotype of Vicia lathyroides was the most asymmetric one. A multivariate analysis using 34 karyological parameters, in addition to the symmetry indices, has been carried out and the corresponding dendrogram of linkage distances showed six different groups. Molecular investigations on the inclusive group in study by employing ITS DNA sequences indicated a different pattern of relationships. The cladistic analysis combining the molecular data set with karyological parameters evidenced that the species of sections Vicia and Atossa join closely to each other in a paraphyletic group, which includes the monophyletic section Wiggersia. Therefore, our karyological and molecular data provide information about the phylogenetic position of the analysed species inside the subgenus Vicia and are discussed in relation to previous results obtained by morphology, isozymes and ribosomal genes analyses.     

Cytological characterization of Vicia oroboides Wulfen in Jacq

Vicia oroboides, a rare taxon belonging to section Atossa of subgenus Vicia, was recovered and analysed by means of cytological and karyological methods with the aim of both characterising this species and integrating our knowledge on phylogeny of subgenus Vicia. Automated karyotype analysis and nuclear DNA content have been determined after Feulgen’s reaction; chromosome banding was performed by fluorochrome staining to evidence heterochromatic blocks along the chromosome complement. The chromosome number is in line with the values of the species of section Atossa; the GC- and AT-rich sites were identified by CMA and DAPI staining. Karyomorphological parameters, based on symmetry indices, provide information about the phylogenetic position of this species inside the subgenus Vicia. DNA content is reported for the first time.     

Cytological and molecular characterization of Vicia esdraelonensis Warb. & Eig: a rare taxon

Summary. Vicia esdraelonensis, a rare taxon belonging to section Hypechusa of subgenus Vicia, was recovered and analyzed by cytological, karyological, and molecular methods, with the aim of both characterizing this species and furthering our knowledge of the phylogeny of subgenus Vicia. Automated karyotype analysis, nuclear DNA content, and chromatin organization were determined by the Feulgen reaction, as well as chromosome banding after double staining with 4′,6-diamidino-2-phenylindole (DAPI) and chromomycin A3. The chromosome number and the nuclear DNA content were in agreement with the values of the species of section Hypechusa. The GC- and AT-rich preferential sites were determined by chromomycin A3 and DAPI staining. Karyomorphological parameters indicated that V. esdraelonensis is in an intermediate position in the spatial representation of the species of section Hypechusa on the basis of symmetry indices, as well as in the dendrogram of linkage distance constructed on 37 chromosome parameters. Molecular data based on internal transcribed spacer sequences show that V. esdraelonensis can doubtlessly be included in section Hypechusa and document its closeness to V. noeana. A cladistic analysis combining the molecular data set with karyological characters is also reported. Karyological, cytological, and molecular data allow characterization of the V. esdraelonensis genome and provide information about the phylogenetic position of this species within the Hyrcanicae series of section Hypechusa. Correspondence and reprints: Dipartimento di Biologia, Università di Pisa, Via L. Ghini 5, 56126 Pisa, Italy.     

Karyological and molecular characterisation of subgenus Vicia (Fabaceae)

In the present report, we have analysed the subgenus Vicia by karyological and molecular approaches with the aim to clarify the relationships among Vicia species included in this subgenus by previously evidenced morphological investigations. Multivariate analysis using several karyomorphological parameters in addition to symmetry indices has allowed the construction of a dendrogram of linkage distances very useful to compare and to include in a phylogenetic tree obtained by internal transcribed spacer (ITS) rDNA sequences. Moreover, a separate analysis was performed combining our molecular data on ITS sequences with those reported in the literature for the section Vicilla. Our analyses partly confirm the monophyletic status of the various sections in which the subgenus Vicia has been divided, however questioning, in some cases, the real need to maintain all the nine sections so far accepted and the placement of some individual species in the two subgenera Vicia and Vicilla.     

Fine characterization of turbellarian chromosomes I. Giemsa and quinacrine banding in Dugesia polychroa (O. Schmidt)

Giemsa and quinacrine banding was routinely produced in metaphase spreads of the freshwater triclad Dugesia polychroa. The techniques reported here may help eliminate the problems in chromosome banding which have prevented the application of differential chromosome banding in karyological studies of this taxon. More detailed karyological and phylogenetic comparisons with other species now seems possible.     

Cytogenetic studies on Brazilian Xyris species (Xyridaceae)

Cytological features are presented for the first time for 12 species and one natural hybrid from 16 Brazilian populations of the genus Xyris , including members of two sections ( Xyris and Nematopus ). The results are compared with a review of all available chromosome numbers previously reported for the genus. All analysed species show semi-reticulate interphase nuclei and prophase to prometaphase chromosomes with heterogeneous condensing behaviour. Chromosome numbers varied between 2 n = 26 and 2 n ≈ 108. In Brazil especially, the karyological differentiation of Xyris seems to be generally characterized by dysploid and polyploid increase of the chromosome number, possibly after interspecific hybridization. This is the first cytological study including species of the section Nematopus and the first karyological investigation including South American species of the genus Xyris . © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 138 , 245–252.     

On the highest chromosome number in mammals

The mitotic and meiotic chromosomes of the semiaquatic rodent Ichthyomys pittieri (Rodentia, Cricetinae) from Venezuela were analyzed by means of conventional staining and several banding techniques. The diploid chromosome number of this rare species is 2n = 92, which is the highest value known for mammals. It is assumed that this exceptionally high chromosome number is the result of repeated centric fissions. The karyotype of I. pittieri was compared with that of Anotomys leander, for which a diploid number of 2n = 92 has also been reported. The karyological relationships existing within the Neotropical Cricetidae are summarized.     

Karyomorphology, heterochromatin patterns and evolution in the genus Ophrys (Orchidaceae)

Karyotype structures and heterochromatin distribution in representative taxa of the genus Ophrys are compared, based on Feulgen-stained and banded somatic metaphase chromosomes. The karyotypes of Ophrys iricolor , O. lupercalis , O. caesiella , O. lutea , O. lunulata , O. x. tardans , O. apifera , O. praecox , O. lacaitae and O. insectifera are described for the first time. The karyological analyses indicate the relationships among the species with respect to asymmetry indices and heterochromatin content. Chromosomal differences have been helpful in clarifying the taxonomic position of Ophrys species that do not have clear affinities. The representative species of Araniferae , Fuciflorae and Ophrys sections exhibited the most asymmetrical karyotypes, while chromosome complements of the O. fusca–O. lutea group, of O. tenthredinifera and of O. bombyliflora proved to be less asymmetrical. Weakly heterochromatic chromosomes, with heterochromatin present mostly in thin centromeric bands, characterize Ophrys C-banded karyotypes. Chromomycin A₃ (CMA) staining revealed that the analysed species exhibit a weak pattern of CMA⁺ bands at centromeric, intercalary or telomeric regions. No DAPI bright blocks were observed. The significance of the karyological data is discussed with regard to the relationships between the analysed species. © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 148 , 87–99.     

Chromosomes of thecercopithecus aethiops species group:C. aethiops (Linnaeus, 1758),C. cynosurus (Scopoli, 1786),C. pygerythrus (Cuvier, 1821), andC. sabaeus (Linnaeus, 1766)

The banded karyotypes of 34 monkeys of known geographic origin and belonging to the Cercopithecus aethiops group of species (C. aethiops, C. pygerythrus, C. cynosurus, C. sabaeus) show that chromosome evolution in this group is highly conservative. All species have 2n =60 chromosomes with very similar chromosome banding. However, differences were found both within and between species. A polymorphism of the NOR area of the “marked” chromosome pairs was found in all taxa (9 of 34 animals). All individuals referred to C. sabaeus,from both West Africa and the Barbados, are characterized by having highly positive G- and C- banded terminal sequences on chromosomes 7,10,12, and 14. Outgroup comparisons with other primates and a parsimony analysis suggest that these terminal bands are derived and are probably good taxonomic and phylogenetic indicators. Moreover, chromosome 18 is variable both between and within species in G banding and in short-arm length. The existence of within-species variation in karyotypes suggests that karyological comparisons must be based on adequate samples that include specimens coming from all the major geographic populations of the species concerned.     

The use of seed proteins revealed by SDS PAGE in taxonomy and phylogeny of some Lathyrus species

Electrophoretic seed protein patterns of 18 samples belonging to 14 species and 4 sections of Lathyrus are treated by principle component analysis (PCA). The morphological ground and karyological structure data of these samples are also discussed in the light of sectional and groups delimitation. The species under study of section Cicercula are separated into 3 groups and L. hirsutus referred to the most primitive species within Lathyrus species. This agrees with their previous grouping delimination based on morphological characters and with chromosomal features such as karyotype structure. L. aphaca referred to the most advanced species in this genus, which agrees with the modification of this morphological characteristics and reduction in chromosome criteria. Section Nissolia hasan intermediate position between section Cicercula and section Aphaca. This revised version was published online in July 2006 with corrections to the Cover Date.     

Cytotaxonomic studies in the sections Spirostachyae (Drejer) Bailey and Ceratocystis Dumort. of the genus Carex L. (Gyperaceae), with special reference to Iberian and North African taxa

LUCEÑO, M. & CASTROVIEJO, S., 1993. Cytotaxonomic studies in the sections Spirostachyae (Dreyer) Bailey and Ceratocystis Dumort of the genus Carex L. (Cyperaceae), with special reference to the Iberian and North African taxa. Karyological studies were carried out on Iberian and North African populations of all species included in sections Spirostachyae and Ceratocystis of the genus Carex , with observations made on meiotic divisions in one to seven individuals per population. In the light of the karyological data we present hypotheses of the most probable phylogeny of the section Spirostachyae , and of the relationships of the sections Spirostachyae, Ceratocystis and Elatae. Carex laevigata and C. camposii are transferred from section Elatae to section Spirostachyae.     

Taxonomic problems in Carthamus (Asteraceae): RAPD markers and sectional classification

Recent molecular studies have clarified the problem of the generic limits of Carthamus . However, taxonomic problems remain in the sectional classification: section Carthamus was confirmed as a natural group, but molecular phylogenies based on DNA sequences did not resolve the other sections, resulting in a confusing conglomerate. A survey based on RAPD markers has been carried out in this conflicting group of sections, with the aim of evaluating the usefulness of these markers at the section and species levels. On the basis of the results, correlated with morphological and karyological characters, some recommendations are made in relation to the taxonomy of Carthamus . The number of sections is reduced from five to two: section Thamnacanthus is currently classified as a different genus, Phonus ; section Carthamus s.s. is retained in its present delimitation, and section Atractylis is redefined, including current sections Atractylis , Lepidopappus and Odontagnathius . The commonly accepted treatment of some taxa as subspecies is not well founded, and C. alexandrinus , C. creticus , C. tenuis and C. turkestanicus should be considered as independent species. Our results confirm that methodological problems posed by RAPD markers can be avoided by careful laboratory procedures and appropriate data analyses; they also suggest that this kind of marker is useful at low taxonomic levels and is, furthermore, complementary to DNA sequence analysis.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 147 , 375–383.     

Analysis of eggplant (<Emphasis Type="Italic">Solanum melongena</Emphasis>)-related germplasm: morphological and AFLP data contribute to phylogenetic interpretations and germplasm utilization

A total of 94 Solanum accessions, including eggplants and related species, were morphologically characterized based on greenhouse observations, and molecularly analysed by the AFLP technique. Morphological parameters were helpful in assessing similarities or differences among accessions, and molecular data were used to support morphological conclusions. A dendrogram was computed based on the Dice genetic distances using the neighbour-joining method. The analysis was efficient in the assignment of a species name for eight out of nine accessions that were not previously classified, and revealed that 14 further accessions were misnamed in the collection originally received. The results indicate that the taxonomy of Solanum sections and subgenera including several species should be reconsidered. The AFLP technique was revealed as an efficient tool in determining genetic relationships among species. In general, morphological observations were consistent with molecular data, indicating that both approaches complemented to define the phylogenetic status of a large genus like Solanum. In terms of eggplant breeding, the molecular analysis of the Melongena complex, and of the other sections of the subgenus Leptostemonum, establishes useful germplasm relationships in the gene pool available for the genetic improvement of the cultivated species. The results we have provided highlight an urgent necessity to include molecular parameters in handling and characterizing the genebank-deposited germplasm related to cultivated crops.Communicated by F. Salamini     

Cytogenetics of the European plethodontid salamanders of the genus Hydromantes (Amphibia,Urodela)

A karyological analysis was carried out on different European species of the genus Hydromantes (Plethodontidae). All the species examined share the same chromosome number (2n=28) and, with the exception represented by pair XIV, morphologically similar karyotypes. While the karyotypes display a similar distribution — mainly centromeric and pericentric — of C-heterochromatin, quantitative variations in pericentric heterochromatin are observed among species. In the continental species Hydromantes italicus and ambrosii as well as in the eastern Sardinian species imperialis, flavus and specie nova, pair XIV consists of heteromorphic sex chromosomes of the XX/XY type. It is proposed that the differentiation of the Y might have taken place through the occurrence of a structural rearrangement, such as a pericentric inversion, starting from a hypothetical, homomorphic pair XIV. A sex-related heteromorphism is not found in the western Sardinian species H. genei. A further karyological differentiation among these species concerns the position of the nucleolus organizing region (NOR), which is located on chromosome XII (H. italicus and ambrosii) or on chromosome X, close to the centromere (H. genei, H. imperialis and H. specie nova), or in an intercalary position (H. flavus). The location and the number of the 5 S DNA sites have been conserved during species divergence. On the basis of these karyological data, as well as of results obtained through a preliminary restriction enzyme analysis of the ribosomal and genomic DNAs, the phyletic relationships among the European Hydromantes species are discussed.     

Phylogenetic relationships in the family Resedaceae L.

A study was made on the phylogenetic relationships of species of the family Resedaceae, based on morphological features, chromosome meiotic behaviour, karyotype features, size and fertility of pollen grains, nucleotypic parameters, seed protein profiles and esterase isozyme patterns.For the comparison of the seed protein profiles among species a method was developed based on the presence or absence of the bands by means of a computer program. The dendrogram obtained by such a method is in line, to a great extent, with the clusters (sections) obtained within the family based on morphological features.Data on meiotic behaviour and on morphology, such as the type of fruit, carpel numbers and others, suggest that x=5 is the primitive basic chromosome number of this family. x₁=6 and x₂=7 are considered as secondary basic numbers derived from x=5 through aneuploid changes.The results support a proposed phylogenetic tree of the genera and sections of the genus Reseda represented in the European Flora.The principal phenomena that have operated in the evolution of the Resedaceae seem to be aneuploid changes, polyploidy and structural rearrangements. A trend towards DNA increase in the evolution of this group is also apparent.     

Multivariate analysis of the Vicia sativa L. aggregate

GIL, J. & CUBERO, J. I., 1993. Multivariate analysis of the Vicia sativa L. aggregate . The sativa group of species was studied by using multivariate analysis and karyological analysis. Thirty biometrical characters were studied by using 92 Vicia sativa L. accessions from a variety of geographical sources. The four main taxa analysed were: sativa, macrocarpa, cordata and angustifolia. Principal component analysis provided clear separation among sativa, cordata and angustifolia in terms of the first two components. Karyotype analysis confirmed the PCA results, and by combining both methods, misclassed specimens can be correctly determined. Cluster analyses using the characters which displayed the greatest loads over the first components were performed in order to identify the most valuable characters from the taxonomic point of view. These were flower length, calyx length, pod width, constriction of the pod between seeds, leaf length, leaflet width, seed length and seed width. Our results suggest an early separation of angustifolia from the ancestral stem. The taxon cordata may be useful in future plant breeding programmes not only for crossing with V. saliva to increase the leaflet size of this taxon, but also as a bridge species between sativa and angustifolia.     

Molecular and karyological aspects of Batoidea (Chondrichthyes, Elasmobranchi) phylogeny

Although considerable progress has been made in elucidating the relationships within the Chondrichthyes, there is no agreement as it concerns the systematics of Batoidea, the most derived superorder among cartilaginous fishes, and many different interpretations exist. Our investigation provides the first assessment of relationships among the described batoid species using sequences from both mtDNA and nuclear genes as well as karyological morphology. Our work consists primarily in reconstructing the phylogenetic relationships of Batoidea by examining the mtDNA (16S) and nuclear gene (18S) sequences from 11 batoid species. The three analytical methods (NJ, MP and Bayesian analysis) grouped Rajiformes, Myliobatiformes and Rhinobatiformes. In these trees the two torpedoes diverge from the other batoid fishes. We also compare the molecular data with the available karyological evidence, which consist of the diploid number and the karyotype morphology of eight species belonging to the four orders examined. The results show that the karyological structure in the different species is generally consistent with the various phylogenetical trees, and that Torpediniformes confirm their unique genome organization.     

HETEROCHROMATIN BANDING IN BOYKINIA,HEUCHERA, MITELLA,SULLIVANTIA, TIARELLA,AND TOLMIEA (SAXIFRAGACEAE)

Karyotypic differences were sought among species of Boykinia, Heuchera, Mitella, Sullivantia, Tiarella, and Tolmiea utilizing a modification of the Hy-banding technique. Prominent centromeric and some telomeric heterochromatin banding was observed. Boykinia aconitifolia and species of Sullivantia possess an identical banded karyotype, while four species of Heuchera, Mitella diphylla, Tiarella cordifolia, and Tolmiea menziesii (the latter at the tetraploid level) are characterized by a second, slightly different banded karyotype. In Sullivantia, Giemsa C-banding stains the same chromosomal regions revealed by Hy-banding. Larger amounts of heterochromatin are present in chromosomes of species of Heuchera, Mitella, Tiarella, and Tolmiea than in chromosomes of Sullivantia species and Boykinia aconitifolia. These karyological observations confirm generic relationships and demonstrate the systematic applicability of chromosome banding techniques to plants with very small chromosomes.     

Karyological studies of Iranian <Emphasis Type="Italic">Allium</Emphasis> L. (Amaryllidaceae) species with focus on sect. <Emphasis Type="Italic">Acanthoprason</Emphasis>. 1. Mitotic chromosomes

Eighteen species and subspecies (34 accessions) of Allium sect. Acanthoprason and 11 species (17 accessions) belonging to other subgenera and sections of Allium were karyologically investigated and include first reports for 12 species. The examined plants of 47 accessions were diploid, three accessions of two species were tetraploid, and in the A. bisotunense accession, we found a mix of di- and triploid individuals. B chromosomes were found in 10 accessions. A basic chromosome number of x = 8 was confirmed for all investigated members of subg. Melanocrommyum and subg. Allium, and x = 9 for Allium tripedale of subg. Nectaroscordum. Idiograms were drawn for each accession, and metaphase images are presented illustrating observed chromosomal variations. Also, karyotype features and asymmetry parameters were calculated for all accessions. Chromosomal aberrations, e.g. aneuploid cells or loss of whole or parts of chromosome arms, were rarely observed. In general, the karyotypes showed low variation in inter- and intrachromosomal asymmetry especially inside of the taxonomic groups, though satellited chromosomes were good markers for subgenera and even specific for two studied sections of subg. Allium. Six different types of satellites were recognized, two of them were newly described: Type P was prevalent in subg. Melanocrommyum, and type O in sect. Codonoprasum. Statistical analyses were performed on five karyological parameters to test correct relationships and also to test previous grouping hypotheses. Although our data confirm distinct karyological characters for the subgenera investigated, the remarkable morphological diversity inside of subg. Melanocrommyum is not mirrored by striking karyological differences.