Chromosome bands in freshwater triclads

Four species of Triclads belonging to three families, Dugesia polychroa and Dugesia mediterranea (Dugesiidae), Planaria torva (Planariidae) and Dendrocoelum lacteum (Dendrocoelidae) were studied for chromosome banding: C-banding for all the species, ASG banding for Dugesia polychroa and G-banding for Dugesia polychroa and Dugesia mediterranea. C-banding results for Dugesia lugubris-polychroa group suggest a high level of heterochromatin evolution in the group. Small bands strictly limited to the centromeric region were seen in Dugusia lugubris and this pattern is similar to the patterns of some species belonging to other families, notably Planaria torva and Dendrocoelum lacteum. This could be considered a primitive character. In contrast numerous heterochromatin bands which are useful to characterize the different karyotypes were seen in Dugesia polychroa and Dugesia mediterranea. Our data suggest that heterochromatin is important in freshwater triclad speciation.     

Analysis of different banding patterns and late replicating regions in chromosomes of Allium cepa,A. sativum and A. nigrum

Different banding procedures and preferential Giemsa staining of late replicating DNA-rich regions were carried out in metaphase chromosomes of three species belonging to different sections of the genus Allium (A. cepa, A. sativum and A. nigrum). The banding, as well as the late replicating patterns were species-specific. The late replicating pattern proved to be, in all cases, the more detailed, and represented the highest percentage of the karyotype differentially stained. Lower percents of the karyotype positively stained were accounted for by C-banding, by modified C-banding and by N-banding. In A. cepa interphase nuclei the pattern of constitutive heterochromatin fitted well with that of late replicating DNA-rich regions, but the coincidence with that revealed by C-banding was only partial. This supports the suggestion that late replicating regions may be considered to be a special category of heterochromatin. On the other hand, it seems that not all C-banded material replicates at the end of the S phase. By the modified C-banding, stained centromere dots or small bands, as well as bands at the NORs are observed.     

Karyomorphological analyses and heterochromatin characteristics disclose phyletic relationships among 2n = 32 and 2n = 36 species ofOrchis (Orchidaceae)

The karyotypes of eight taxa ofOrchis L. with 2n = 32 and 2n = 36 have been investigated using morphometrical measurements following staining with Feulgen, Giemsa (C-banding) and the DNA specific fluorochrome Hoechst 33258. The karyotypes ofO. coriophora subsp.fragrans, andO. papilionacea proved to be the most asymmetrical, whileO. morio andO. longicornu exhibited the most symmetrical karyotypes. Using C-banding and the fluorochrome H33258 only the taxa with high asymmetry indices showed the presence of differentially stained chromatin bands. In most chromosomes heterochromatin bands were present at the telomeric position. The present results seem to indicate that the analysed species do not form a homogeneous group and further subdivisions are possible, which, in turn, do not always correlate with divisions based on morphological characters. Both karyomorphology and heterochromatin distribution coincide in indicating a possible evolutionary pathway.     

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 meaning of DAPI bands observed after C-banding and FISH procedures

Abstract

Under specific technical conditions chromosome staining with 4′,6-diamidino-2-phenylindole (DAPI) permits characterization of heterochromatic regions as AT-rich (DAPI⁺) or AT-poor (DAPI^?), especially when the chromosomes are counterstained with chromomycin A₃ (CMA), which preferentially binds to GC-rich DNA. DAPI⁺ bands also often have been observed after C-banding or FISH. In these cases, however, it is not clear whether only AT-rich regions stain positively with DAPI or other heterochromatins with different base compositions also are stained. We evaluated the meaning of DAPI bands observed after C-banding and FISH using three plant species bearing different types of heterochromatin: DAPI⁺/CMA^?, DAP^?/CMA⁺ and DAPI⁰/CMA⁰ (neutral bands). Additional tests were performed using propidium iodide, a fluorochrome without preferential affinity for AT or GC. Our results indicate that AT-rich heterochromatin stains as DAPI⁺ bands after C-banding or FISH, but other kinds of heterochromatin also may be stained by DAPI.     

Cytogenetic study of F1 hybrids betweenCrepis dinarica andCrepis froelichiana (Asteraceae)

Crepis dinarica andC. froelichiana are two closely related species of theC. praemorsa complex. Even though they exhibit the same chromosome number (2n = 8) and similar idiogram shape, they differ widely in quantity and distribution of heterochromatin bands. The hybrids between these two species comprise three morphological types. Parental genomes were distinguished in hybrids by Giemsa differential staining (C-banding). Although meiosis presents only a few abnormalities (about 2.4%), the percentage of aborted pollen grains is very high (90%).     

GIEMSA C-BANDED KARYOTYPES OF SEVEN NORTH AMERICAN SPECIES OF ALLIUM

The karyotypes of seven North American Allium species were studied by Giemsa C-banding technique. Two species (A. shoenoprasum and A. tricoccum) were diploids with 2n = 16 chromosomes. Three species (A. cernuum, A. douglasii and A. geyeri) were also diploids but with 2n = 14 chromsomes. Diploid and tetraploid populations of A. textile (2n = 14, 28) were studied. The population of A. canadense studied here was a tetraploid (2n = 28). All these North American species, except A. geyeri, possessed centromeric bands in all their chromosomes and nucleolar constriction bands in their satellited chromosomes. Allium shoenoprasum contained telomeric bands in most of its chromosomes but the other species had them only in a small number of chromsomes. Only three species (A. shoenoprasum, A. textile and A. tricoccum) were found to have intercalary bands in some chromosomes. The heterochromatin distribution in B chromosomes of three species was also observed. In A. cernuum, the heterochromatin occupied most of the length of all its Bs, but in A. shoenoprasum, heterochromatin was concentrated in the centromeric region. One population of A. textile (CC1179) was found to have a B chromosome in which very little heterochromatin existed.     

X-linked heterochromatin distribution in the holocentric chromosomes of the green apple aphid <Emphasis Type="Italic">Aphis pomi</Emphasis>

Chromatin organization in the holocentric chromosomes of the green apple aphid Aphis pomi has been investigated at a cytological level after C-banding, NOR, Giemsa, fluorochrome staining and fluorescent in situ hybridization (FISH). C-banding technique showed that heterochromatic bands are exclusively located on X chromosomes. This data represents a peculiar feature that clearly contradicts the equilocal distribution of heterochromatin typical of monocentric chromosomes. Moreover, silver staining and FISH carried out with a 28S rDNA probe localized rDNA genes on one telomere of each X chromosome; CMA₃ staining reveals that these silver positive telomeres are the only GC-rich regions among A. pomi heterochromatin, whereas all other C-positive bands are DAPI positive thus containing AT-rich DNA.     

Cold-sensitive chromosome regions and heterochromatin inCestrum aurantiacum (Solanaceae)

The karyotype ofCestrum aurantiacum was analyzed for the presence of coldsensitive regions (CSRs) and other types of constitutive heterochromatin. A range of techniques was employed including the fluorescent DAPI, chromomycin/DAPI double staining and actinomycin D/DAPI counter-staining, and the non-fluorescent C-banding applied as single or sequential staining, sequential N-banding and silver impregnation. Four classes of constitutive heterochromatin were recognized: CSRs, nucleolar organizers, non-nucleolar chromomycin-positive bands, and indifferently fluorescent bands. The banded karyotype ofC. aurantiacum is compared with those of otherCestrum species. The sectionsHabrothamnus andCestrum are not karyologically distinct.     

Karyomorphological parameters and C-band distribution suggest phyletic relationships within the subtribeLimodorinae (Orchidaceae)

Karyotype attributes and heterochromatin distribution were used to characterize fourteen taxa of the subtribeLimodorinae (Orchidaceae). The karyotypes were established using morphometrical parameters following Feulgen staining and C-banding. No significant differences in heterochromatin content were found between specimens collected from various sites. Four species of theEpipactis helleborine group possess some chromosome pairs with quite similar heterochromatin patterns; some differences were found inE. distans with respect to other species of this group.Epipactis palustris differed significantly from otherEpipactis species in its different karyotype and its numerous terminal C-bands. The largest differences from the other genera were shown inLimodorum as far as karyomorphology and heterochromatin patterns were concerned. C-band distribution indicated similarity among non-homologous chromosomes, supporting a possible palaeo-polyploid origin for theCephalanthera andEpipactis karyotypes.     

Chromosome evolution in Australian rodents

The chromosome complements of 188 specimens of 29 species of Australian murid rodents belonging to the subfamilies Pseudomyinae and Hydromyinae and the Uromys/Melomys group have been compared. At least one specimen of 18 different species was successfully C-banded. — The autosomal complements of many (9) diverse Pseudomyinae, one species of Melomys and one Hydromyinae proved to be identical, comprising 48 elements in the diploid set, the two smallest autosomal pairs of which are metacentric. No other karyotype is common to more than one species. From this we conclude that these three groups have been derived from a common ancestor which also possessed such a karyotype. The genus Zyzomys is exceptional since it possesses only 44 elements and lacks the two smallest metacentrics. — Karyotypic evolution within this apparently single phyletic line has been remarkably conservative, only three rearrangements being required to derive the most divergent karyotype. Moreover most of the observed rearrangements involve pericentric inversions and only one example of a fusion was found. Considerable differences in heterochromatin content, as determined by C-banding, occur between species however. Two species proved exceptional in this respect, namely Notomys cervinus and Uromys caudimaculatus. N. cervinus possesses numerous heterochromatic short arms. In U. caudimaculatus, there is a striking difference between northern and southern populations; in the former heterochromatin is present principally in the telomeric areas of the conventional A-chromosomes whereas in the latter it is found as separate supernumerary chromosomes. — In contrast to the autosomes, the X and Y chromosomes show high inter- and intra-specific variability in both size and morphology. All of this variability can be explained in terms of variation in heterochromatin content. Moreover the amount of heterochromatin in the X and Y chromosomes is highly correlated both within and between species. The Y chromosome of Uromys caudimaculatus is, however, distinctive in that it lacks C-banding.     

Cold-sensitive chromosome regions and heterochromatin inCestrum (Solanaceae):C. strigillatum,C. fasciculatum,andC. elegans

Cold-induced mitotic under-condensation of certain chromosome segments is a rare phenomenon in plants. There are about 11 genera of monocotyledons and only 3 of dicotyledons, where species are known to have such cold-sensitive regions (CSRs). The molecular causes of cold-induced undercondensation are not clear, and no consistent cytochemical characteristics of CSRs are known. Recently we have presented a chromosome banding analysis on CSRs and their relation to constitutive heterochromatin inCestrum parqui (Solanaceae), a species of sect.Cestrum. The present study is concerned with a similar analysis inC. strigillatum of sect.Cestrum, and inC. fasciculatum andC. elegans of sect.Habrothamnus. Chromomycin/DAPI fluorescent double staining, sequential C-banding, and sequential silver impregnation were applied. The species differ in detail but are similar qualitatively. Four classes of heterochromatin can be discriminated. (1) CSRs, with banding properties indicating AT-rich constitutive heterochromatin. After cold-treatment CSR heterochromatin can be silver-impregnated from interphase, as chromocentres, to metaphase, as undercondensed segments. CSRs are subject to frequent heteromorphy. (2) Nucleolar organizers. Two pairs were identified in the karyotypes. Banding properties indicate GC-rich heterochromatin. The nucleolar organizing regions are less evident and their silver-reducing capability reduces during metaphase. (3) Non-nucleolar CMA-positively fluorescing bands. These are minute, polymorphic, positively C-stained, and restricted to one or a few sites in the karyotypes. (4) Indifferently fluorescing, positively C-stained bands. They occur on centromeres, some chromosome ends, and clustered over the chromosome arms. They are mostly very delicate and do not resist harsh banding treatments. — The species investigated here andC. parqui resemble each other qualitatively in heterochromatin classes (1), (2), and (3), but differ much in banding properties of class (4). Therefore, heterochromatin characteristics in the genus are not so uniform as the present results inC. strigillatum, C. fasciculatum, andC. elegans appear to show.     

Two types of constitutive heterochromatin in the chromosomes of some Fritillaria species

A Giemsa banding technique has been used to study C-banding in mitotic chromosomes in root tips of Fritillaria graeca, F. crassifolia and F. rhodocanakis, all diploids (2n=24) belonging to the graeca group. In the first two the C-bands were of two types, diverging in respect of staining regularly and specifically within chromosomes. In one type it was weak, being intermediate between that of intensely stained ones, representing the other class, and the euchromatin. In F. graeca the pale bands were proximally localized and confined to 5 pairs, whereas in F. crassifolia they occurred only in the 4 M chromosomes, in each within the centromeric constriction as a large inclusion. The interphase nuclei of both species contained pale and heavily stained chromocentres. No pale ones occurred in such nuclei of F. rhodocanakis. The probability is discussed that the two classes of C-band represent distinct types of heterochromatin, differing both in respect of condensation throughout the whole mitotic cycle and in the repetitive DNA sequences they most likely contain. In all 3 species pairs of Giemsa-positive centromeric dots, representing the centromeres, were masked both by proximally or centromerically localized bands, irrespective of the class of heterochromatin they represented.     

Giemsa C-banded karyotypes in Serupius L. (Orchidaceae)

Giemsa C-banding is utilized for the first time to characterize eight taxa of the genus Serapias . Heterochromatin distribution indicated that the Serapias species form a very homogeneous group. All the species possess chromosome pairs with similar heterochromatin patterns. C-banding showed conspicuous bands located around the centromeres, with some het-erochromatic short arms. There was more heterochromatin in S. apulica and S. nurrika than in the other taxa. Extensive centromeric heterochromatin may indicate recent structural rearrangements in the chromosome complement. Taken altogether, karyomorphology indicates a rather recent origin for the genus Serapias , which might also account for the small amount of interspecific variation observed.     

Heterochromatin and chromosome variation in cultivated species ofTulipa subg.Eriostemones (Liliaceae)

The chromosomes of several cultivatedTulipa species belonging to the subg.Eriostemones were examined using conventional staining and C-banding techniques. Most of the species have lightly banded chromosomes with heterochromatin content varying from nil to about 15%. The banding patterns of several taxa are described and discussed in regard to species relationships.     

Cytogenetics of the Brazilian Bolitoglossa paraensis (Unterstein, 1930) salamanders (Caudata,Plethodontidae)

Plethodontid salamanders of genus Bolitoglossa constitute the largest and most diverse group of salamanders, including around 20% of living caudate species. Recent studies have indicated the occurrence of five recognized species in the Brazilian Amazon Rainforest. We present here the first cytogenetic data of a Brazilian salamander, which may prove to be a useful by contribution to the cytotaxonomy of the genus. Specimens were collected near the “type” locality (Utinga, Belém, PA, Brazil). Chromosomal preparations from duodenal epithelial cells and testes were subjected to Giemsa staining, C-banding and DAPI/CMA₃ fluorochrome staining. All specimens showed a karyotype with 13 bi-armed chromosome pairs (2n = 26). Nucleolar Organizer Regions, evidenced by CMA₃, were located distally on the long arm of pair 7 (7q). DAPI+ heterochromatin was predominantly centromeric, with some small pericentromeric bands. Although the C-banding patterns of other Bolitoglossa species are so far unknown, cytogenetic studies conducted in other Plethodontid salamanders have demonstrated that pericentromeric heterochromatin is a useful cytological marker for identifying interspecific homeologies. Species diversification is usually accompanied by chromosomal changes. Therefore, the cytogenetic characterization of Bolitoglossa populations from the middle and western Brazilian Amazon Basin could identify differences which may lead to the identification of new species.     

Comparison of the chromosomes of Triticum timopheevi with related wheats using the techniques of C-banding and in situ hybridization

Summary The chromosomes of the tetraploid wheats Triticum timopheevi (Genome AAGG) and T. araraticum (Genome AAGG) were C-banded at mitosis. The identity of the banded and unbanded chromosomes was then established by firstly making comparisons with the hexaploid species T. zhukovskyi which has the genome formula AAAAGG. Secondly, the meiotic pairing in F₁ hybrids between T. timopheevi and diploid wheats was examined by means of C-banding. The results showed that the banded chromosomes belonged to the G genome, while the unbanded chromosomes belonged to the A genome. Only one of the two pairs of satellited chromosomes had strong heterochromatic bands. The relationship between the genomes of T. timopheevi and T. dicoccum (Genome AABB) was then assessed at meiosis in hybrids between these species, using the techniques of C-banding and in situ hybridisation of a cloned ribosomal RNA gene probe. It was concluded that there were differences both in the amount and distribution of heterochromatin and also translocation differences between the species.     

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.     

Chromosomes of antelope squirrels (genus Ammospermophilus): A systematic banding analysis of four species with unusual constitutive heterochromatin

The G- and C-banding patterns of mitotic chromosomes from four species of antelope squirrels (Ammospermophilus harrisi, interpres, leucurus and nelsoni) are discussed with special attention payed to the unusual quantities and position of constitutive heterochromatin. Heterochromatin appears to be responsible for the observation that cells from antelope squirrels contain over 70% more DNA than cells from another ground squirrel. A substantial fraction of this excess DNA consists of sequences that band as satellites in neutral CsCl or Cs₂SO₄-Ag⁺ density gradients. Interspecies similarities in the distribution of heterochromatin suggest that it has a function of some importance to these species and has therefore been conserved.     

Comparison of the Giemsa C-banded karyotypes of Hystrix coreana and H. patula (Poaceae, Triticeae)

The karyotypes of Hystrix coreana from eastern USSR and H. patula from USA were investigated by Giemsa C-banding. Both species are outbreeders and have 2n = 4x = 28. The karyotype of two plants of H. coreana has 10 metacentric, 6 submetacentric, 8 heterobrachial and 4 SAT chromosomes; two plants differed by having 12 metacentric, 4 submetacentric, 8 heterobrachial and 4 SAT-chromosomes, and 10 metacentric, 4 submetacentric, 9 heterobrachial and 5 SAT-chromosomes, respectively. The C-banding pattern had no or few inconspicuous intercalary bands, but conspicuous telomeric C-bands in one or both arms giving a high content of heterochromatin (16.3–18.2%). The chromosome complement of one plant of H. patula had 8 metacentric, 6 submetacentric, 8 heterobrachial and 6 SAT-chromosomes. The C-banding pattern had between 1 and 4 intercalary or centromeric bands and conspicuous telomeric bands on one or both arms giving a high content of constitutive heterochromatin (16.4%).