Comparison of C-banding patterns and in situ hybridization sites using highly repetitive and total genomic rye DNA probes of `Imperial' rye chromosomes added to `Chinese Spring' wheat

1992 ◽  
Vol 67 (2) ◽  
pp. 71-83
Author(s):  
Yasuhiko MUKAI ◽  
Bernd FRIEBE ◽  
Bikram S. GILL
Genome ◽  
1994 ◽  
Vol 37 (3) ◽  
pp. 477-481 ◽  
Author(s):  
Jie Xu ◽  
R. L. Conner ◽  
A. Laroche

'Agrotana', a wheat-alien hybrid (2n = 56), is a potential source of resistance to common root rot, stem rust, wheat streak mosaic virus, and the wheat curl mite. However, the origin of 'Agrotana', reported to be durum wheat × Agropyron trichophorum (pubescent wheatgrass), is uncertain. The objective of this investigation was to determine the chromosome constitution of 'Agrotana' using C-banding and fluorescence in situ hybridization techniques. The F1 hybrid of 'Agrotana' × 'Chinese Spring' wheat showed 7 I + 21 II in 14.9% of the pollen mother cells, evidence of the presence of the A, B, and D genomes in 'Agrotana'. The hybrid had 16 heavily C-banded chromosomes, namely 4A, and 1-7B of wheat, and a translocation that probably involved wheat chromosomes 2A and 2D. In situ hybridization using biotinylated genomic DNA of Ag. trichophorum cv. Greenleaf blocked with CS DNA failed to identify the alien chromosomes in 'Agrotana', indicating that the alien chromosomes were not likely derived from pubescent wheatgrass. In situ hybridization using labelled wheat genomic DNA blocked with 'Agrotana' DNA revealed that 'Agrotana' had 40 wheat, 14 alien, and 2 (a pair) wheat–alien translocated chromosomes. There was no homology between wheat and the alien chromosomes or chromosome segments involved in the wheat–alien recombinant. Two of the seven pairs of alien chromosomes were homoeologous to each other. The ability to identify alien chromatin in wheat using labelled wheat DNA instead of labelled alien DNA will be particularly useful in chromosome engineering of wheat germplasms having alien chromatin of unknown origin.Key words: wheat–alien hybrid, C-banding, fluorescence in situ hybridization, labelled wheat DNA as probe.


1983 ◽  
Vol 25 (4) ◽  
pp. 319-323 ◽  
Author(s):  
J. Hutchinson ◽  
T. E. Miller ◽  
S. M. Reader

The meiotic chromosome pairing of one 'Chinese Spring' wheat aneuhaploid (3AL) and two 'Chinese Spring' aneuploid × rye hybrids (N3A-T3B and N5B-T5D × rye) were studied. These genotypes all display higher than normal levels of pairing which were analysed with the aid of the technique of C-banding. The results show that the pattern of pairing is neither random nor of an unrestricted homoeologous nature. Pairing is more frequent between unhanded (A and D genome) chromosomes, than between either unhanded (A or D genome) and banded (B or R genome) chromosomes, or between banded (B and R genome) chromosomes.


1985 ◽  
Vol 27 (5) ◽  
pp. 549-558 ◽  
Author(s):  
Rama S. Kota ◽  
Jan Dvořák

A rapid technique is described for the production of a disomic substitution of an alien chromosome for a selected wheat chromosome directly from the amphiploid. Determination of homoeology between the chromosomes involved is a by-product of this technique. The technique involves the production of a nullisomic amphiploid from a cross between a wheat mono-telosomic and a diploid species and recurrently backcrossing it as a male twice to the monotelosomic. Disomic and ditelosomic substitutions are then selected upon self-pollination. The utility of this technique is shown by producing a large number of disomic and ditelosomic substitutions of an Aegilops longissima chromosome homoeologous to wheat chromosome 6B from two populations of Ae. longissima. The incorporated Ae. longissima chromosome was characterized by C-banding and in situ hybridization of an 18S–26S rRNA gene probe. The chromosome differs in C-band pattern from chromosome 6B of 'Chinese Spring' and does not pair with the 6Bp telosome at metaphase I. It also differs from 'Chinese Spring' chromosome 6B by having the 18S–26S rRNA compound locus in the long arm.Key words: disomic substitution, homoeology, C-banding, in-situ hybridization, rRNA genes, gene synteny, Triticum, Aegilops longissima.


Genome ◽  
1995 ◽  
Vol 38 (3) ◽  
pp. 435-442 ◽  
Author(s):  
A. Cabrera ◽  
B. Friebe ◽  
J. Jiang ◽  
B. S. Gill

C-banding patterns of Hordeum chilense and of Triticum aestivum 'Chinese Spring' – H. chilense disomic addition lines were analyzed and compared with in situ hybridization patterns using a biotin-labeled highly repetitive Triticum tauschii DNA sequence, pAs1, and a wheat 18S–26S rDNA probe. All seven H. chilense chromosomes pairs and the added H. chilense chromosomes present in the addition lines were identified by their characteristic C-banding pattern. Chromosome morphology and banding patterns were similar to those of the corresponding chromosomes present in the parent H. chilense accession. A C-banded karyotype of the added H. chilense chromosomes was constructed and chromosome lengths, arm ratios, and relative length, as compared with chromosome 3B, were determined. The probe pAs1 was found to hybridize to specific areas on telomeres and interstitial sites along the chromosomes, allowing the identification of all seven pairs of the H. chilense chromosomes. Comparison of the patterns of distribution of the hybridization sites of clone pAs1 in the T. tauschii and H. chilense chromosomes was carried out by in situ hybridization on somatic metaphase chromosomes of the HchHchDD amphiploid. In situ hybridization using the 18S–26S rDNA probe confirmed that the H. chilense chromosomes 5Hch and 6Hch were carrying nucleolus organizer regions. The results are discussed on the basis of phylogenetic relationships between D and Hch genomes.Key words: Hordeum, Triticum, C-banding, in situ hybridization, phylogeny.


Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1819
Author(s):  
Tatyana Karamysheva ◽  
Svetlana Romanenko ◽  
Alexey Makunin ◽  
Marija Rajičić ◽  
Alexey Bogdanov ◽  
...  

The gene composition, function and evolution of B-chromosomes (Bs) have been actively discussed in recent years. However, the additional genomic elements are still enigmatic. One of Bs mysteries is their spatial organization in the interphase nucleus. It is known that heterochromatic compartments are not randomly localized in a nucleus. The purpose of this work was to study the organization and three-dimensional spatial arrangement of Bs in the interphase nucleus. Using microdissection of Bs and autosome centromeric heterochromatic regions of the yellow-necked mouse (Apodemus flavicollis) we obtained DNA probes for further two-dimensional (2D)- and three-dimensional (3D)- fluorescence in situ hybridization (FISH) studies. Simultaneous in situ hybridization of obtained here B-specific DNA probes and autosomal C-positive pericentromeric region-specific probes further corroborated the previously stated hypothesis about the pseudoautosomal origin of the additional chromosomes of this species. Analysis of the spatial organization of the Bs demonstrated the peripheral location of B-specific chromatin within the interphase nucleus and feasible contact with the nuclear envelope (similarly to pericentromeric regions of autosomes and sex chromosomes). It is assumed that such interaction is essential for the regulation of nuclear architecture. It also points out that Bs may follow the same mechanism as sex chromosomes to avoid a meiotic checkpoint.


2006 ◽  
Vol 72 (8) ◽  
pp. 5311-5317 ◽  
Author(s):  
Kengo Kubota ◽  
Akiyoshi Ohashi ◽  
Hiroyuki Imachi ◽  
Hideki Harada

ABSTRACT Low signal intensity due to poor probe hybridization efficiency is one of the major drawbacks of rRNA-targeted in situ hybridization. There are two major factors affecting the hybridization efficiency: probe accessibility and affinity to the targeted rRNA molecules. In this study, we demonstrate remarkable improvement in in situ hybridization efficiency by applying locked-nucleic-acid (LNA)-incorporated oligodeoxynucleotide probes (LNA/DNA probes) without compromising specificity. Fluorescently labeled LNA/DNA probes with two to four LNA substitutions exhibited strong fluorescence intensities equal to or greater than that of probe Eub338, although these probes did not show bright signals when they were synthesized as DNA probes; for example, the fluorescence intensity of probe Eco468 increased by 22-fold after three LNA bases were substituted for DNA bases. Dissociation profiles of the probes revealed that the dissociation temperature was directly related to the number of LNA substitutions and the fluorescence intensity. These results suggest that the introduction of LNA residues in DNA probes will be a useful approach for effectively enhancing probe hybridization efficiency.


1991 ◽  
Vol 40 (1) ◽  
pp. 117-120 ◽  
Author(s):  
Avirachan T. Tharapel ◽  
Mazin B. Qumsiyeh ◽  
Paula R. Martens ◽  
Sugandhi A. Tharapel ◽  
James D. Dalton ◽  
...  

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