Karyotyping Melandrium album, a dioecious plant with heteromorphic sex chromosomes

Genome ◽  
1990 ◽  
Vol 33 (4) ◽  
pp. 556-562 ◽  
Author(s):  
D. D. Ciupercescu ◽  
J. Veuskens ◽  
A. Mouras ◽  
D. Ye ◽  
M. Briquet ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Sex Chromosomes ◽  
Nucleolus Organizer ◽  
Rrna Genes ◽  
Hybridization Technique ◽  
Metaphase Chromosomes ◽  
Nucleolus Organizer Regions ◽  
Melandrium Album ◽  
Group B

Mitotic metaphase chromosomes of Melandrium album obtained from root protoplasts were studied. Morphologically, the chromosomes were either metacentrics or submetacentrics. They were classified into three distinct groups: group A comprising six pairs of autosomal metacentrics, group B comprising five pairs of autosomal submetacentrics, and the sex chromosomes: X and Y. The X chromosome is a metacentric (r = 1.44), which accounts for more than 14% of the genome. The Y chromosome is a metacentric with, virtually, equal arms (r = 1.09) and accounts for 21% of the genome, being the largest of the complement. The Y:X ratio was 1.4. Ethidium bromide, caffeine, and vinblastine were used to obtain a better resolution and higher frequency of satellited chromosomes 7q and 9p. The proposed karyotype of M. album is 2n = 24, XX, s(7q;9p) for female and 2n = 24, XY, s(7q;9p) for male plants. Nucleolus organizer regions (NORs) were present at the telomeric sites of three chromosome pairs: 7q, 9p, and 10p. The NORs were polymorphic, particularly between the nonhomologous chromosomes. The in situ hybridization technique localized the rRNA genes on four chromosome pairs: 5p, 7q, 9p, and 10p. The discrepancy between the NORs and the hybridization signals was probably due to the fact that NORs were restricted only to transcriptionally active rRNA genes. It was concluded that for a complete description and characterization of rRNA genes, both NOR detection and in situ hybridization techniques, as complementary methods, should be employed.Key words: Melandrium album, karyotype, satellites, idiogram, nucleolus organizer regions, in situ hybridization.

scholarly journals Co-amplification of rRNA genes with CAD genes in N-(phosphonacetyl)-L-aspartate-resistant Syrian hamster cells.

1983 ◽  
Vol 3 (11) ◽  
pp. 2066-2075 ◽  
Author(s):  
G M Wahl ◽  
L Vitto ◽  
J Rubnitz
Keyword(s):  
In Situ Hybridization ◽  
Syrian Hamster ◽  
Blot Hybridization ◽  
Nucleolus Organizer ◽  
Rrna Genes ◽  
28S Rrna ◽  
Metaphase Chromosomes ◽  
Rdna Genes ◽  
Resistant Mutants

The amplified CAD genes in N-(phosphonacetyl)-L-aspartate (PALA)-resistant Syrian hamster cells are located in an expanded chromosomal region emanating from the site of the wild-type gene at the tip of the short arm of chromosome B-9. The terminus of B-9 in PALA-sensitive cells contains a cluster of rRNA genes (i.e., a nucleolus organizer, rDNA). We have used a molecular clone containing sequences complementary to Syrian hamster 28S rRNA to investigate whether rDNA is coamplified with CAD genes in the PALA-resistant mutants. In situ hybridization of this probe to metaphase chromosomes demonstrates that rDNA and CAD genes do coamplify in two independently isolated PALA-resistant mutants. The tight linkage of CAD and rDNA genes was demonstrated by their coordinate translocation from B-9 to the end of the long arm of chromosome C-11 in one mutant. Blot hybridization studies substantiate the in situ hybridization results. Both types of analysis indicate that only one or two rDNA genes, on the average, are coamplified per CAD gene. The data are consistent with the model that unequal exchanges between rDNA genes mediate the amplification of CAD genes in the Syrian hamster mutants that were analyzed.

Co-amplification of rRNA genes with CAD genes in N-(phosphonacetyl)-L-aspartate-resistant Syrian hamster cells

1983 ◽  
Vol 3 (11) ◽  
pp. 2066-2075
Author(s):  
G M Wahl ◽  
L Vitto ◽  
J Rubnitz
Keyword(s):  
In Situ Hybridization ◽  
Syrian Hamster ◽  
Blot Hybridization ◽  
Nucleolus Organizer ◽  
Rrna Genes ◽  
28S Rrna ◽  
Metaphase Chromosomes ◽  
Rdna Genes ◽  
Resistant Mutants

The amplified CAD genes in N-(phosphonacetyl)-L-aspartate (PALA)-resistant Syrian hamster cells are located in an expanded chromosomal region emanating from the site of the wild-type gene at the tip of the short arm of chromosome B-9. The terminus of B-9 in PALA-sensitive cells contains a cluster of rRNA genes (i.e., a nucleolus organizer, rDNA). We have used a molecular clone containing sequences complementary to Syrian hamster 28S rRNA to investigate whether rDNA is coamplified with CAD genes in the PALA-resistant mutants. In situ hybridization of this probe to metaphase chromosomes demonstrates that rDNA and CAD genes do coamplify in two independently isolated PALA-resistant mutants. The tight linkage of CAD and rDNA genes was demonstrated by their coordinate translocation from B-9 to the end of the long arm of chromosome C-11 in one mutant. Blot hybridization studies substantiate the in situ hybridization results. Both types of analysis indicate that only one or two rDNA genes, on the average, are coamplified per CAD gene. The data are consistent with the model that unequal exchanges between rDNA genes mediate the amplification of CAD genes in the Syrian hamster mutants that were analyzed.

Single-copy and amplified CAD genes in Syrian hamster chromosomes localized by a highly sensitive method for in situ hybridization

1982 ◽  
Vol 2 (3) ◽  
pp. 308-319
Author(s):  
G M Wahl ◽  
L Vitto ◽  
R A Padgett ◽  
G R Stark
Keyword(s):  
In Situ Hybridization ◽  
Syrian Hamster ◽  
Specific Radioactivity ◽  
Large Chromosome ◽  
Hybridization Technique ◽  
Wild Type ◽  
Aspartate Transcarbamylase ◽  
Metaphase Chromosomes ◽  
Cad Gene

Syrian hamster cells resistant to N-(phosphonacetyl)-L-aspartate (PALA), a specific inhibitor of the aspartate transcarbamylase activity of the multifunctional protein CAD, overproduce this protein as a result of amplification of the CAD gene. We have used a sensitive in situ hybridization technique to localize CAD genomes in spreads of metaphase chromosomes from several independent PALA-resistant lines and from wild-type PALA-sensitive cells. The amplified genes were always found within chromosomes, usually in an expanded region of the short arm of chromosome B9. In wild-type cells, the CAD gene was also on the short arm of chromosome B9. In one mutant line, 90 to 100 CAD genes were found within an expanded B9 chromosome and 10 to 15 more were near the distal end of one arm of several different chromosomes. Another line contained most the genes in a telomeric chromosome or large chromosome fragment. The amplified genes were in chromosomal regions that were stained in a banded pattern by trypsin-Giemsa. A few double minute chromosomes were observed in a very small fraction of the total spreads examined. The it situ hybridizations were performed in the presence of 10% dextral sulfate 500, which increases the signal by as much as 100-fold. Using recombinant DNA plasmids nick-translated with [125I]dCTP to high specific radioactivity, 10 CAD genes in a single chromosomal region were revealed after 1 week of autoradiographic exposure, and the position of the unique gene could be seen after 1 month.

scholarly journals Single-copy and amplified CAD genes in Syrian hamster chromosomes localized by a highly sensitive method for in situ hybridization.

1982 ◽  
Vol 2 (3) ◽  
pp. 308-319 ◽  
Author(s):  
G M Wahl ◽  
L Vitto ◽  
R A Padgett ◽  
G R Stark
Keyword(s):  
In Situ Hybridization ◽  
Syrian Hamster ◽  
Specific Radioactivity ◽  
Large Chromosome ◽  
Hybridization Technique ◽  
Wild Type ◽  
Aspartate Transcarbamylase ◽  
Metaphase Chromosomes ◽  
Cad Gene

Syrian hamster cells resistant to N-(phosphonacetyl)-L-aspartate (PALA), a specific inhibitor of the aspartate transcarbamylase activity of the multifunctional protein CAD, overproduce this protein as a result of amplification of the CAD gene. We have used a sensitive in situ hybridization technique to localize CAD genomes in spreads of metaphase chromosomes from several independent PALA-resistant lines and from wild-type PALA-sensitive cells. The amplified genes were always found within chromosomes, usually in an expanded region of the short arm of chromosome B9. In wild-type cells, the CAD gene was also on the short arm of chromosome B9. In one mutant line, 90 to 100 CAD genes were found within an expanded B9 chromosome and 10 to 15 more were near the distal end of one arm of several different chromosomes. Another line contained most the genes in a telomeric chromosome or large chromosome fragment. The amplified genes were in chromosomal regions that were stained in a banded pattern by trypsin-Giemsa. A few double minute chromosomes were observed in a very small fraction of the total spreads examined. The it situ hybridizations were performed in the presence of 10% dextral sulfate 500, which increases the signal by as much as 100-fold. Using recombinant DNA plasmids nick-translated with [125I]dCTP to high specific radioactivity, 10 CAD genes in a single chromosomal region were revealed after 1 week of autoradiographic exposure, and the position of the unique gene could be seen after 1 month.

Primer-induced in situ hybridization to plant chromosomes

Genome ◽  
1993 ◽  
Vol 36 (5) ◽  
pp. 815-817 ◽  
Author(s):  
S. Abbo ◽  
T. E. Miller ◽  
I. P. King
Keyword(s):  
In Situ Hybridization ◽  
Mitotic Chromosome ◽  
Organizer Region ◽  
Nucleolus Organizer Region ◽  
Nucleolus Organizer ◽  
Hybridization Technique ◽  
Plant Chromosomes ◽  
Secale Cereale L ◽  
Future Potential

Primer-induced in situ hybridization of high copy sequences was used successfully on rye mitotic chromosome spreads. Nucleolus organizer sequences were detected in rye, and the pattern obtained was in full agreement with previously reported results by conventional in situ hybridization. The future potential of the primer-induced in situ hybridization technique is briefly discussed.Key words: primer-induced in situ hybridization, rye (Secale cereale L.), nucleolus organizer region.

Molecular cytogenetics of Icelandic birch species: physical mapping by in situ hybridization and rDNA polymorphism

1995 ◽  
Vol 25 (1) ◽  
pp. 101-108 ◽  
Author(s):  
Kesara Anamthawat-Jonsson ◽  
J.S. Heslop-Harrison
Keyword(s):  
In Situ Hybridization ◽  
Physical Mapping ◽  
Molecular Cytogenetics ◽  
Ribosomal Gene ◽  
Nucleolus Organizer ◽  
Environmental Conservation ◽  
Nucleolus Organizer Regions ◽  
A Genome ◽  
Breeding Programmes

The physical mapping of genes can reveal the organization of a genome and identify relationships of plant species, especially where they are involved in interspecific hybridization and polyploidy. Here we determine the chromosomal locations of the major ribosomal gene family (18S-5.8S-26S rDNA) by fluorescent in situ hybridization in two Icelandic birch species, Betulapubescens Ehrh. and Betulanana L. In the tetraploid birch (B. pubescens), the rDNA was localized on four major and two minor sites, while the diploid dwarf birch (B. nana) had four major sites. The major loci in both species were in nucleolus organizer regions, close to the centromeres of a pair of metacentric and a pair of sub-metacentric chromosomes. The dispersed interphase in situ hybridization pattern showed gene expression at all major sites. The two additional loci in B. pubescens, when detected, appeared to be sub-telomeric and inactive at interphase. Southern analysis of rDNA showed considerable restriction fragment length polymorphism in B. pubescens. Some polymorphism may reflect gene flow among populations and between the two co-existing birch species. The understanding of genome relationships, gene introgression, and evolution of birch species will be important to the breeding programmes steered towards environmental conservation and forestry.

The Relationship between the (In-)Stability of NORs and Their Chromosomal Location: The Example of Cercopithecidae and a Short Review of Other Primates

2017 ◽  
Vol 153 (3) ◽  
pp. 138-146 ◽  
Author(s):  
Michèle Gerbault-Seureau ◽  
Lauriane Cacheux ◽  
Bernard Dutrillaux
Keyword(s):  
Chromosome Evolution ◽  
Short Review ◽  
Nucleolus Organizer ◽  
Fragile Sites ◽  
Metaphase Chromosomes ◽  
Nucleolus Organizer Regions ◽  
Size Number ◽  
Almost All ◽  
The Relationship

Amongst Cercopithecidae, the species of the Cercopithecini tribe underwent a very active chromosome evolution, principally by fissions, which increased their chromosome number up to 72. In contrast, all the species of Papionini have fairly similar karyotypes with 42 chromosomes. In animals, nucleolus organizer regions (NORs) are generally considered as instable structures, which frequently vary in size, number, and location at both infra- and interspecific levels. Although in Cercopithecinae the NORs, involved in breaks, exchanges, and translocations, behave like fragile sites in somatic cells, their number and location appear to be very stable between species. Fluorescence in situ hybridization of a 28S rDNA probe on metaphase chromosomes displayed a unique interstitial location in either an acrocentric pair (in 12 species of Cercopithecini) or a metacentric pair (in 6 species of Papionini). A non-exhaustive survey of literature data on NOR location in other primates shows that numerical variations of the NORs principally depend on their location: most multiple NORs are in terminal positions, while almost all unique NORs are in interstitial positions. We propose that this correlation is the consequence of the selection against gametic imbalances involving the chromosomal material distal to the NORs, which is effective when they are interstitially, but not terminally, located. Thus, the consequences of the interstitial NOR instability for reproduction are essentially limited to their size variations, as observed in Cercopithecidae.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

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