Karyotype analysis and heterochromatin differentiation with Giemsa C-banding and fluorescent counterstaining inCephalanthera (Orchidaceae)

Detailed studies of the chromosomes of the three Austrian species of the genusCephalanthera showed them all to have basically similar karyotypes. BothC. damasonium (2n = 36) andC. longifolia (2n = 32) have three large and several classes of smaller chromosome pairs. The karyotype ofC. rubra (2n = 44) is composed of four large and several groups of smaller pairs. The heterochromatin in these species amounts to about 10% of total karyotype length. All the chromosomes have Giemsa-positive centromeres, but only a few have intercalary or terminal bands. Using differential fluorescent staining with DAPI/actinomycin D, quinacrine/actinomycin D (both A-T specific), and chromomycin A₃/distamycin A (G-C specific) three different types of major heterochromatic bands can be characterized in respect of their satellite DNA composition: highly A-T rich, slightly A-T rich, and very G-C rich. The chromosomes ofC. longifolia contain more A-T rich C-bands than those ofC. damasonium, while the latter's have more G-C rich heterochromatin. In both species several C-bands appear as secondary constrictions or gaps in the Feulgen-stained chromosomes, but most likely, in each species there is only one pair of chromosomes where the secondary constrictions function as nucleolus organizing regions. No major intraspecific variation could be observed except on one small chromosome pair ofC. longifolia which had a heteromorphic C-band in most individuals. Possible pathways of karyotype evolution involving polyploidy and Robertsonian events are discussed.     

Giemsa C-banded karyotypes of some diploidArtemisia species

Detailed C-banded karyotypes of eight diploidArtemisia species from three different sections are reported together with preliminary observations on three additional related diploid species. In the majority, the overall amount of banding is relatively low. Bands are mostly confined to distal chromosome regions; intercalary banding is virtually absent and centromeric heterochromatin is also scarce. With the exception ofA. judaica there is in general great uniformity in karyotype structure but considerable interspecific variation in total karyotype length (and hence DNA content) ranging from 44 µm inA. capillaris (2n = 18) to 99 µm inA. atrata (2n = 18).A. judaica (2n = 16; total karyotype length 97 µm) was distinguished by its karyomorphology, with one large non-banded metacentric chromosome pair and 7 pairs of smaller terminally banded meta- or submetacentric chromosomes.     

Giemsa C-banded karyotypes inCapsicum (Solanaceae)

Giemsa C-banding is applied for the first time inCapsicum, allowing preliminary karyotype differentiation of six diploid species. Comparison of interphase nuclei and heterochromatic C-bands reveals striking differences between taxa and contributes to their taxonomic grouping. Therefore, C-banding appears to be a powerful tool for the cytogenetics and karyosystematics of the genus. Banding patterns are characterized by the omnipresence of centromeric bands and a variable number of smaller to larger distal bands, with the addition of intercalary bands in some cases. Satellites are always C-positive. Relationships between species and possible trends of karyotype evolution are discussed, with special reference to the origin of x = 13 from x = 12 and the increase of heterochromatin, regarded as advanced features.Chromosome studies inCapsicum (Solanaceae), III. For the first and the second part seeMoscone (1990, 1993).     

The karyotype ofFestucopsis serpentini (Poaceae Triticeae) from Albania studied by banding techniques and in situ hybridization

The karyotypes of two populations ofFestucopsis serpentini (2n = 2x = 14) endemic to Albania were investigated in detail by Giemsa C- and N-banding, AgNO₃ staining, and in situ hybridization with an rDNA probe. The complements consisted of 14 large chromosomes, 10 metacentric and 4 SAT-chromosomes, a metacentric and a submetacentric pair. SAT-chromosomes from one population carried exclusively minute satellites, whereas SAT-chromosomes from another population also carried larger polymorphic satellites, suggesting a geographical differentiation. The existence of four chromosomes with nucleolus forming activity was established through AgNO₃ staining; however, the rDNA probe additionally hybridized to intercalary positions in the short arms of two metacentric chromosomes revealing two inactive rDNA sites. C-banding patterns comprised from zero and up to four very small to larger, generally telomeric bands per chromosome giving low levels of constitutive heterochromatin. Similarities in chromosome morphology and C-banding patterns identified the homologous relationships of all chromosomes in one population, but of three pairs only in the other. Reliable identification of homologous chromosomes between plants was only possible for the SAT-chromosomes. A comparison between the C-banded karyotypes ofF. serpentini andPeridictyon sanctum supports their position in two genera.     

A survey of C-band patterns in chromosomes ofLilium (Liliaceae)

C-band patterns are described for 20Lilium spp. distributed across six sections. All species have a similar basic karyotype (n = 12) but C-bands differ markedly between them. The patterns are characterized by a dispersed scattering of thin intercalary bands as well as centric and NOR bands. Only one species,L. canadense, shows a clear equilocal pattern with intercalary C-bands occurring proximally in all of the longer chromosome arms. Comparing species, similar patterns are revealed forL. regale andL. sulphureum, forL. formosanum andL. longiflorum (all in sect.Leucolirion) and to a lesser extent forL. hansonii, L. martagon, andL. tsingtauense (sect.Martagon). The pattern forL. henryi (previously classed in sect.Sinomartagon) matches those ofL. regale andL. sulphureum quite well and its transfer to sect.Leucolirion is proposed. This is consistent with results from interspecies hybrids betweenL. henryi andL. regale (and related species) which are reportedly fertile. No other clear similarities in C-band patterns were seen across species. It seems that C-band patterns change rapidly inLilium and hence their usefulness in classification will be restricted to identifying closely related species.Dedicated to Prof.D. G. Catcheside on the 80th anniversary of his birth.     

Analysis of nucleolus organizer regions (NORs) in mitotic and polytene chromosomes ofPhaseolus coccineus by silver staining and Giemsa C-banding

Chromosomes with active nucleolus organizer regions (NORs) were visualized in root tip metaphases ofPhaseolus coccineus using the silver staining technique. A mean number of 5.5 Ag-NORs per cell was observed in 54 cells from eight plants. In the endopolyploid nuclei of the suspensor the silver technique did not demonstrate the reported specificity for nucleolus organizer activity, because there was usually pale staining of nucleoli and preferential staining of heterochromatic regions in the polytene chromosomes including pericentromeric material, telomeres and NORs. The mean number of NORs per nucleolus as detected by this method was 5.8 (28 nucleoli analysed). Using a modified preparation technique, giant chromosomes stained pale, but nucleoli of suspensor cells displayed darkly silver staining internal domains, each of which originating from a nucleolus organizer.—Giemsa C-banding of endopolyploid suspensor nuclei revealed C-positive nucleolus organizers with darkly staining intranucleolar fibrils. The latter were frequently involved in inter-NOR associations. In 34 nucleoli analysed, the mean number of Giemsa C-positive NORs per nucleolus was 6.0.Dedicated to Professor Dr.Lothar Geitler on the occasion of his 80th birthday.     

Cytotaxonomy and karyotype evolution inPhleum sect.Phleum (Poaceae) in Poland

Three taxa are distinguished in the sectionPhleum Griseb. in Poland:P. nodosum (2n = 14),P. pratense (2n = 42), and the third one unformally named hereP. commutatum (2n = 14). It corresponds morphologically toP. commutatum Gaud. reported as a tetraploid taxon (2n = 28) from other geographic regions. Giemsa C-banded karyotypes of these three taxa help clarify the taxonomic status ofP. commutatum and the origin of the hexaploidP. pratense. It is suggested that changes in the amount of telomeric heterochromatin played an important role in the evolution ofPhleum karyotypes.     

Heterochromatin and silver banding of rye (Secale cereale,Gramineae) chromosomes

Air-dried chromosomes of rye when stained with aqueous silver nitrate show differential banding patterns. In addition to staining the NOR sites, the silver nitrate stains all regions of constitutive heterochromatin, as identified by Giemsa C-banding, as well as a number of small interstitial regions. However, the heterochromatin on the B chromosome is not stained by the silver method. This is proposed as a rapid and reliable banding method.     

Infraspecific variation in C-banded karyotype and chiasma frequency inCucumis sativus (Cucurbitaceae)

Infraspecific cytogenetical variation was studied in a diverse collection of five non-cultivated and cultivatedCucumis sativus accessions. The individual chromosomes of different accessions could be identified by the C-banding pattern and chromosome measurements. About 40–50% of the genomic area are made up of heterochromatin inC. sativus. The non-cultivated accessions exhibit more heterochromatin and lower chiasma frequencies per pollen mother cell than cultivated accessions. There is infraspecific variation in C-banding pattern, karyomorphology and multinucleolate cells. The use of C-banding in infraspecific classification is discussed.     

Contribution to the orchid flora of South Central Africa

11 new taxa (8 new species, 3 new varieties) ofOrchidaceae are described from South Central Africa, including data on their habitat and distribution and discussions of their systematic affinities.     

Giemsa banded karyotypes,systematics, and evolution inAnacyclus (Asteraceae-Anthemideae)

Giemsa C-banding allows the differentiation of six, otherwise very similar karyotypes from the small genusAnacyclus. Banding style—with stable centromeric and nucleolar bands, and diverse specific banding patterns in distal chromosome segments—contributes significantly to generic demarcation and systematic grouping. The amount of banding corresponds to heterochromatic chromocentres and increases from perennials to annuals. Relationships with other nucleotype parameters and evolutionary mechanisms are discussed.First contribution of a series on Giemsa Banded Karyotypes, Systematics, and Evolution inAnthemideae (Asteraceae).     

Karyosystematic studies on threeCrepis species (Asteraceae) endemic to Greece

This work examines the cytogeographical distribution, the morphological characters, and the karyotypes of threeCrepis species endemic to Greece (C. sibthorpiana, C. incana, andC. heldreichiana). C. sibthorpiana is diploid (2n = 2x = 8),C. incana is diploid (2n = 2x = 8) and tetraploid (2n = 4x = 16, 17), andC. heldreichiana is always dekaploid (2n = 10x = 40). The Giemsa positive bands, usually pairs of dots, are mainly centromeric inC. incana, while they are terminal inC. sibthorpiana (on the short arm of all chromosomes) and inC. heldreichiana (on both arms of all chromosomes). Intercalary C-bands are scarce and usually variable within karyotypes, individuals, and species. The most variable karyotype both in Feulgen and Giemsa preparations is that ofC. incana, in which also supernumerary chromosomes were observed, which are polysomic to standard set members. On the basis of morphological and karyological data the evolutionary relationships among the threeCrepis taxa are discussed.     

Chromosome banding patterns in Lettuce species (Lactuca sect.Lactuca,Compositae)

Chromosome banding patterns obtained with C- and N- banding, and AgNO₃ staining were studied in somatic metaphase complements of fourLactuca species.L. sativa andL. serriola have almost identical chromosome morphology, andL. saligna differs only slightly from them, butL. virosa is quite distinct from the other species. A gross comparison of the banded karyotypes suggests a closer relationship ofL. saligna toL. sativa/serriola than toL. virosa. Our data agree with the results of previous crossing experiments in these species but conflict partly with recent RFLP data which indicate a closer phenetic relationship ofL. saligna toL. virosa than toL. sativa/serriola. Such a discrepancy may be explained assuming that domestication ofL. sativa/serriola resulted in an increased selection pressure on unique DNA sequences as demonstrated by the RFLP data. Differential evolution of specific heterochromatin classes (and presumably of highly repetitive DNA classes), as revealed by chromosome banding techniques was not linked to domestication. Thus the disparity in conclusions about relationship (in terms of genetic similarity) as based on the different experimental approaches reflects a non-parallel evolution of highly repetitive vs. unique DNA classes.     

Giemsa C-banded karyotypes ofAllium ericetorum andA. kermesinum (Alliaceae)

The widely distributedAllium ericetorum and the local endemic of the Steiner Alps (Slovenia),A. kermesinum, are two closely related species of sect.Rhizirideum, whose main distinguishing character is perianth colour. To obtain further evidence for species separation, karyotype morphology and C-banding patterns were examined in 10 populations. The chromosome number was 2n = 16. In some populations ofA. ericetorum a B-chromosome occurred. Arm and satellite lengths and C-banding patterns were subjected to cluster analysis. Three different karyotype classes were observed and described. Karyotypes did not clearly discriminate between plants with different colours of perianth segments and therefore did not provide evidence for a taxonomic separation ofA. ericetorum and A. kermesinum. There is polymorphism in number and patterns of C-bands within the populations. No correlation between B-chromosomes and particular banding patterns was observed.     

DNA contents,giemsa banding,and systematics inScilla bifolia,S. drunensis,andS. vindobonensis (Liliaceae)

DNA contents have been determined cytophotometrically in the three Central European, relatedScilla speciesS. bifolia (2n = 18, 2 x, 1 C = 6.2 pg),S. drunensis (2n = 36, 4 x, 1 C = 12.8 pg), andS. vindobonensis (2n = 18, 2 x, 1 C = 9.4 pg). The tetraploid speciesS. drunensis contains twice as much DNA as the diploidS. bifolia. However, the diploid speciesS. vindobonensis differs in DNA content fromS. bifolia by a factor of about 1.5. This difference is largely due to euchromatic DNA, although the higher DNA content inS. vindobonensis is combined with higher heterochromatin content. The data indicate thatS. bifolia andS. drunensis on the one hand, andS. vindobonensis on the other hand are phyletically well separated. Previous taxonomic conclusions from morphology as well as C-banding are thus corroborated.Evolution ofScilla and Related Genera, V.     

Giemsa C-banded karyotypes of two subspecies ofHordeum brevisubulatum from China

C-banding patterns ofH. brevisubulatum subsp.brevisubulatum (2x) and subsp.turkestanicum (4x) had conspicuous telomeric C-bands in at least one chromosome arm with a minor difference in average band size between subspecies. Other conspicuous bands were few in number as in other taxa of the species complex. The C-banded area of the chromosomes was estimated to be 7 to 8 and 6 per cent, respectively. C-banding- and SAT-chromosome polymorphisms were observed in both subspecies. The latter and previous observations indicate that the number of SAT-chromosomes is a less reliable diagnostic character. Nucleolar organizer region polymorphisms were demonstrated through silver nitrate staining of nucleoli. C-banding patterns corroborated that tetra- and hexaploid cytotypes of subsp.turkestanicum form an autopolyploid series. Reliable identification ofH. brevisubulatum taxa based on cytological criteria should include the simultaneous use of C-banding patterns, and number and morphology of marker chromosomes.     

Giemsa C-banded karyotypes and taxonomic relationships of some North AmericanAllium species and their relationship to Old World species (Liliaceae)

The somatic karyotypes of six North AmericanAllium species and the EuropeanA. scorzonerifolium have been investigated using a Giemsa C-banding technique. All species have a chromosome number of 2n = 14. InA. scorzonerifolium and the three North American speciesA. dichlamydeum, A. fibrillum andA. unifolium C-bands are restricted to two pairs of nucleolar chromosomes. Each chromosome has a band proximal to the nucleolar constriction and a positively banded satellite. InA. acuminatum, in addition to the bands associated with the nucleolar constrictions, all chromosomes also have pericentromeric bands.A. cernuum exhibits a distinctive banding style: two chromosome pairs with bands adjacent to the nucleolar constrictions and four pairs with telomeric bands on their short arms. In the karyotype ofA. geyeri neither C-bands nor nucleolar chromosomes were found.—A comparison of the banding styles together with other cytological and morphological characters of these species with old world members ofAllium reveals:A. cernuum closely resembles species within subgenusRhizirideum, whereas the other species studies exhibit many similarities with subgenusMolium. Their sectional grouping and their relationships with Old World species are discussed.     

Characterization of different types of condensed chromatin inCostus (Zingiberaceae)

The chromatin structure of six diploids species ofCostus was analysed using conventional Giemsa staining, C-banding and DAPI/CMA fluorochromes. The interphase nuclei in all the species show an areticulate structure and the prophase chromosomes show large blocks of proximal condensed chromatin. After banding procedures, each chromosome exhibits only centromeric dot-like DAPI⁺/CMA^– C-bands whereas the satellites (one pair at each karyotype) are weakly stained after C-banding and show a DAPI^–/CMA⁺ fluorescence. Two chromocentres show bright fluorescence with CMA and weak staining after C-banding whereas the others chromocentres show only a small fraction of DAPI⁺ heterochromatin. These results were interpreted to mean that the greater part of the condensed chromatin has an euchromatic nature whereas two types of well localized heterochromatin occur in a small proportion. The Z-stage analysis suggests that heterochromatin and condensed euchromatin decondense at different times. The chromosome number and morphology of all species are given and the implications of the condensed euchromatin are discussed.Dedicated to Prof.Elisabeth Tschermak-Woess on the occasion of her 70th birthday.     

Interspecific relationship of ten European orchid species as revealed by enzyme electrophoresis

Ten species of orchid plants belonging to the generaOrchis (7),Dactylorhiza (2), andGymnadenia (1) were analyzed by enzyme electrophoresis. Each species can be identified by a combination of enzyme bands different from those of all other species examined. The electrophoretic data were used for the construction of phenetic and phylogenetic trees with the help of computer programs. The trees were almost identical regardless which method was used. Our results differ considerably from a classification based on morphological evidence. The electrophoretic data indicate that the genusOrchis is not a monophyletic group.     

Cytological identification of the genomes in pentaploidAllium neapolitanum using Giemsa C-banding

Two homoeologous sets of chromosomes in pentaploidAllium neapolitanum Cyr. (Liliaceae) are recognizable by their C-bands. The banding pattern is the same for one Californian and five Yugoslavian populations, suggesting a common chromosomal origin. Predominant meiotic association of identically banded homologues indicates a genomic formula of AA A_I BB, confirms an allopolyploid origin, and argues against genetically controlled pairing.