scholarly journals Chromosome Banding in Amphibia. XXXIII. Demonstration of 5-Methylcytosine-Rich Heterochromatin in Anura

2016 ◽  
Vol 148 (1) ◽  
pp. 35-43
Author(s):  
Michael Schmid ◽  
Claus Steinlein

An experimental approach using monoclonal anti-5-methylcytosine (5-MeC) antibodies and indirect immunofluorescence was elaborated for detecting 5-MeC-rich chromosome regions in anuran chromosomes. This technique was applied to mitotic metaphases of 6 neotropical frog species belonging to 6 genera and 4 families. The hypermethylation patterns were compared with a variety of banding patterns obtained by conventional banding techniques. The hypermethylated DNA sequences are species-specific and located exclusively in constitutive heterochromatin. They are found in centromeric, pericentromeric, telomeric, and interstitial positions of the chromosomes and adjacent to nucleolus organizer regions. 5-MeC-rich DNA sequences can be embedded both in AT- and GC-rich repetitive DNA. The experimental parameters that have major influence on the reproducibility and quality of the anti-5-MeC antibody labeling are discussed.

2019 ◽  
Vol 157 (1-2) ◽  
pp. 53-64 ◽  
Author(s):  
Michael Schmid ◽  
Claus Steinlein ◽  
Alina M. Reiter ◽  
Michail Rovatsos ◽  
Marie Altmanová ◽  
...  

An experimental approach using monoclonal anti-5-methylcytosine antibodies and indirect immunofluorescence was elaborated for detecting 5-methylcytosine-rich chromosome regions in reptilian chromosomes. This technique was applied to conventionally prepared mitotic metaphases of 2 turtle species and 12 squamate species from 8 families. The hypermethylation patterns were compared with C-banding patterns obtained by conventional banding techniques. The hypermethylated DNA sequences are species-specific and are located in constitutive heterochromatin. They are highly reproducible and often found in centromeric, pericentromeric, and interstitial positions of the chromosomes. Heterochromatic regions in differentiated sex chromosomes are particularly hypermethylated.


2017 ◽  
Vol 152 (1) ◽  
pp. 38-45 ◽  
Author(s):  
Michael Schmid ◽  
Claus Steinlein ◽  
Christian Lomb ◽  
Marianne Volleth

5-Methylcytosine-rich heterochromatic regions were demonstrated in metaphase chromosomes of 5 species of Chiroptera by indirect immunofluorescence using a monoclonal anti-5-methylcytosine antibody. These species belong to 4 genera and 2 families and are characterized by divergent karyotypes. One species (Glauconycteris beatrix) has an extremely low diploid chromosome number of 2n = 22 with only meta- to submetacentric elements and remarkably large amounts of constitutive heterochromatin located in the centromeric and pericentromeric regions of all chromosome pairs. Two species (G. beatrix and Neoromicia cf. guineensis) possess X-autosome translocations. In all species, the hypermethylated chromosome segments correspond to constitutive heterochromatin, and the numbers and positions of hypermethylated chromosome segments in the karyotypes are constant and species-specific. In some species (Pipistrellus hesperidus, Neoromicia cf. somalicus), there are several smaller chromosome pairs in which the bright anti-5-methylcytosine antibody labeling is not restricted to constitutively heterochromatic regions but is observed along the whole lengths of these chromosomes. The nature of these additional hypermethylated regions is discussed. The analysis of 5-methylcytosine-rich chromosome regions elucidates valuable data for chiropteran cytogenetics and reflects the high pace of evolution of the repetitive DNA fraction in their genomes.


Genome ◽  
1990 ◽  
Vol 33 (4) ◽  
pp. 542-555 ◽  
Author(s):  
B. Friebe ◽  
N.-S. Kim ◽  
J. Kuspira ◽  
B. S. Gill

Cytogenetic studies in Triticum monococcum (2n = 2x = 14) are nonexistent. To initiate such investigations in this species, a series of primary trisomics was generated from autotriploids derived from crosses between induced autotetraploids and diploids. All trisomics differed phenotypically from their diploid progenitors. Only two of the seven possible primary trisomic types produced distinct morphological features on the basis of which they could be distinguished. The chromosomes in the karyotype were morphologically very similar and could not be unequivocally identified using standard techniques. Therefore, C-banding was used to identify the chromosomes and trisomics of this species. Ag–NOR staining and in situ hybridization, using rDNA probes, were used to substantiate these identifications. A comparison of the C-banding patterns of the chromosomes of T. monococcum with those of the A genome in Triticum aestivum permitted identification of five of its chromosomes, viz., 1A, 2A, 3A, 5A, and 7A. The two remaining chromosomes possessed C-banding patterns that were not equivalent to those of any of the chromosomes in the A genome of the polyploid wheats. When one of these undesignated chromosomes from T. monococcum var. boeoticum was substituted for chromosome 4A of Triticum turgidum, it compensated well phenotypically and therefore genetically for the loss of this chromosome in the recipient species. Because this T. monococcum chromosome appeared to be homoeologous to the group 4 chromosomes of polyploid wheats, it was designated 4A. By the process of elimination the second undesignated chromosome in T. monococcum must be 6A. Analysis of the trisomics obtained led to the following conclusions. (i) Trisomics for chromosome 3A were not found among the trisomic lines analyzed cytologically. (ii) Primary trisomics for chromosomes 2A, 4A, 6A, and 7A were positively identified. (iii) Trisomics for the SAT chromosomes 1A and 5A were positively identified in some cases and not in others because of polymorphism in the telomeric C-band of the short arm of chromosome 1A. (iv) Trisomics for chromosome 7A were identified on the basis of their distinct phenotype, viz., the small narrow heads and small narrow leaves. Because rRNA hybridizes lightly to nucleolus organizer regions on chromosome 1A and heavily to nucleolus organizer regions on chromosome 5A, our results indicate that trisomics in line 50 carry chromosome 1A in triple dose and trisomics in lines 28 and 51 carry chromosome 5A in triplicate. Variable hybridization of the rDNA probe to nucleolus organizer regions on chromosomes in triple dose in lines 7, 20, and 28 precluded the identification of the extra chromosome in these lines. Cytogenetic methods for unequivocally identifying trisomics for chromosomes 1A and 5A are discussed. Thus six of the series of primary trisomics have been identified. Telotrisomic lines are also being produced.Key words: Triticum monococcum, trisomics, C-banding, Ag-NOR staining, in situ hybridization, rDNA probes, plant morphology.


Genome ◽  
1991 ◽  
Vol 34 (4) ◽  
pp. 638-643 ◽  
Author(s):  
J. P. M. Camacho ◽  
J. Cabrero ◽  
E. Viseras ◽  
M. D. Lopez-Leon ◽  
J. Navas-Castillo ◽  
...  

A G banding technique combining trypsin and hot saline treatments was used to analyze the chromosomes of two grasshopper species, Eyprepocnemis plorans and Locusta migratoria, both of which contain both standard and supernumerary heterochromatin. Although this technique does not produce G bands like those in mammalian chromosomes, it serves to characterize heterochromatic regions whose nature has been inferred from other banding techniques (C, N, CMA, and DAPI banding). The light regions revealed by G banding contain GC-rich DNA sequences, the more prominent of which coincide with nucleolus organizer regions (NORs). Furthermore, the proximal heterochromatin in E. plorans was heterogeneous, and the standard and supernumerary heterochromatin showed conspicuous differences in organization. Supernumerary heterochromatin is an exception to the regular patterns shown by the standard heterochromatin. The findings are related to the mechanism of action of these banding techniques.Key words: banding techniques, grasshoppers, Eyprepocnemis plorans, Locusta migratoria.


1985 ◽  
Vol 27 (6) ◽  
pp. 665-682 ◽  
Author(s):  
B. Mayr ◽  
D. Schweizer ◽  
M. Mendelak ◽  
J. Krutzler ◽  
W. Schleger ◽  
...  

Chromomycin A3 banding of the mitotic sets of 10 species of Bovidac (cattle, wisent, yak, banteng, gaur, red buffalo, swamp buffalo, sheep, mufflon, and goat) serves to demarcate both centromeric constitutive heterochromatin and R-banding patterns capable of identifying all the chromosomes within a given complement. In all species significant amounts of chromomycin-bright heterochromatin are present at the centromeres of all autosomes, though there was a high degree of intra- and inter-individual variation in the size of the heterochromatic blocks. Marked interspecies differences in the centromeric patterns were evident. The X chromosomes contained appreciable amounts of centromeric heterochromatin only in the two buffaloes. All the animals studied lacked distamycin A – diamidinophenylindole type heterochromatin. AgNO3 staining was applied sequentially to detect the location of active nucleolus organizer regions (NORs). The distribution of NORs was reasonably conservative in most of the species. An exceptional situation was found in the two buffaloes, where only one NOR pair matched with the standard karyotype of the Bovidae.Key words: heterochromatin, chromomycin A3 fluorescence, nucleolus organizers, Bovidae.


1992 ◽  
Vol 43 (5) ◽  
pp. 889 ◽  
Author(s):  
CE May ◽  
R Appels

We have used the techniques of molecular genetics to analyse the nucleolus organizer regions (Nor loci) of a broad range of Australian, Chinese, and other hexaploid wheats (Triticum aestivum L. em Thell.). Genomic DNA was extracted from the leaves of 260 wheat cultivars, 94 different F1 hybrids, and from more than 800 F2 plants. Samples of the DNA were digested with the restriction enzyme TaqI and electrophoretically separated in agarose gels. The DNA fragments were then transferred to DNA filters by Southern blotting and assayed with the probe pTa250.4. This probe specifically detects the intergenic spacer DNA sequences which alternate with the ribosomal-RNA (rRNA) genes in highly repeated tandem arrays within the Nor loci. Analyses of the spacer DNA restriction fragments present in P, F1 and F2 plants show that allelic variants of the Nor loci can be distinguished by the existence of different lengths of spacer DNA fragments, different combinations of different length fragments, or differing amounts of the same length fragments. Eight alleles of the Nor-B1 locus on chromosome 1B (Nor-Bla to Nor-B1h) and 20 variants of the Nor-B2 locus on chromosome 6B (Nor-B2a to Nor-B2t) have been identified. The DNA fragments present in the different alleles are expressed in a co-dominant fashion to give phenotypic F2 ratios of 1 : 2 : 1 and 1 : 2 : 1 : 2 : 4 : 2 : 1 : 2 : 1, indicating segregation at one or two loci, respectively. With two exceptions, individual alleles segregated in a 1 : 1 ratio. On the basis of the presence of different combinations of Nor-B1 and Nor-B2 alleles, the wheat cultivars investigated were divided into 53 phenotypic groups. These groups clearly reflect the germplasm used in wheat breeding programs in different countries. Hybrid wheats could also be recognized and plants with new combinations of alleles were produced.


Genome ◽  
1992 ◽  
Vol 35 (3) ◽  
pp. 436-442 ◽  
Author(s):  
Kurt Weising ◽  
Dieter Kaemmer ◽  
Franz Weigand ◽  
Jörg T. Epplen ◽  
Gunter Kahl

Synthetic oligonucleotides complementary to simple repetitive DNA sequences were used to detect inter- and intra-specific polymorphisms in a leguminous crop plant (chickpea, Cicer arietinum) and its wild relatives. All the investigated repetitive motifs [(GACA)4, (GATA)4, (GTG)5, (CA)8, (TCC)5, (GGAT)4, and (AGTTT)4] were abundantly present and polymorphic in the chickpea genome. Different probes revealed different levels of variability. Whereas species-specific banding patterns were obtained with the (GTG)5 probe, other probes revealed differences between accessions, or even individuals. The somatic multilocus patterns were stable for all probes.Key words: genetic polymorphism, simple repetitive sequences, DNA fingerprinting, synthetic oligonucleotide probes.


1979 ◽  
Vol 21 (1) ◽  
pp. 1-8 ◽  
Author(s):  
L. M. Henderson ◽  
A. N. Bruère

There are ten nucleolus organizer regions (NORs) in domestic sheep (Ovis aries L.), cattle (Bos taurus L.), goat (Capra hircus L.) and aoudad (Ammotragus lervia Blyth) and these are located terminally on chromosomes with homologous G-banding patterns. The similarity in number of nucleolus organizer regions in these species may indicate that their ribosomal DNA regions are infrequently involved in exchange events which could lead to different numbers of active nucleolus organizer regions. Other possible explanations of the conservation of number of nucleolus organizer regions in these species are discussed. The homology of NOR location in these species supports the idea that the Bovidae karyotype tends to be fairly stable apart from changes due to centric fusion events.


1984 ◽  
Vol 26 (4) ◽  
pp. 436-444 ◽  
Author(s):  
I. Linde-Laursen ◽  
R. von Bothmer

In the hybrid Hordeum vulgare × Psathyrostachys fragilis the two genomes were differentiated (i) by length, the P. fragilis chromosomes being 31% longer than the H. vulgare chromosomes; (ii) by a difference in staining intensity of C-banded chromosomes (of possible use for exact localization of breakpoints), the H. vulgare chromosomes being the more heavily stained; (iii) by widely different C-banding patterns; and (iv) by the difference between N-banded H. vulgare and non-N-banded P. fragilis chromosomes. Only C-banding patterns identified each chromosome. Aneuploid cells had lost between one and five P. fragilis chromosomes. Loss of H. vulgare chromosomes is ascribed to squashing. No haploid H. vulgare cell was observed. The P. fragilis chromosomes were characterized by diminished centric constrictions, suppression of nucleolar constrictions, and nucleolus activity, i.e., differential amphiplasty, and generally a peripheral location on the metaphase plate. The same characteristics are normally observed in hybrids producing haploids H. vulgare, suggesting a common mechanism of chromosome elimination. Some cells had a side-by-side arrangement of genomes. The only effect of the hybrid condition on H. vulgare chromosomes was the formation of wider nucleolar constrictions and larger nucleolus organizer regions (NORs) than in parental H. vulgare, suggesting a compensational mechanism for nucleolus activity. The passage of H. vulgare chromosomes through the hybrid to the dihaploid did not influence chromosomal characteristics.Key words: Hordeum, Psathyrostachys, hybrids, elimination of chromosomes, banding.


1998 ◽  
Vol 21 (1) ◽  
pp. 77-80 ◽  
Author(s):  
A. Gava ◽  
T.R.O. Freitas ◽  
J. Olimpio

Intraspecific karyotype variation in mammal species is very common and often caused by centromeric fusion of acrocentric chromosomes. We describe here a new karyotype 2n = 62 (FN = 112) for the genus Cavia from the Moleques do Sul Islands, of the southern coast of Brazil. We analyzed two male and four female karyotypes that had twenty-four biarmed pairs and six pairs of acrocentric chromosomes. The sexual pair consisted of a metacentric X-chromosome and a large acrocentric Y. C-bands were found in the centromeric and pericentromeric regions of almost all chromosomes, except for some small biarmed and acrocentric ones. Nucleolus organizer regions appeared in two biarmed chromosomes, and G-banding patterns were also seen.


Sign in / Sign up

Export Citation Format

Share Document