Comparison of karyotype and rDNA-distribution in somatic chromosomes of Bowenia species (Stangeriaceae, Cycadales)

Somatic chromosomes at mitotic metaphase of Bowenia serrulata, B. spectabilis and B. sp. ‘Tinaroo’ is investigated by the standard aceto-orcein staining method and the fluorescent in situ hybridisation method (FISH) with ribosomal DNA (rDNA) probe. Bowenia serrulata, B. spectabilis and B. sp. ‘Tinaroo’ each have a chromosome number of 2n = 18. The karyotype of B. serrulata exhibits 10 median-centromeric chromosomes, while B. spectabilis and B. sp. ‘Tinaroo’ exhibit eight median-centromeric chromosomes. By using FISH, B. serrulata, B. spectabilis and B. sp. ‘Tinaroo’ show a hybridisation signal on the satellite of the short arm of two submedian-centromeric chromosomes. However, the other hybridisation signal pattern is different among B. serrulata, B. spectabilis and B. sp. ‘Tinaroo’.

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Genome structure and evolution in the allohexaploid weed Avena fatua L. (Poaceae)

Genome ◽

10.1139/g98-154 ◽

1999 ◽

Vol 42 (3) ◽

pp. 512-518 ◽

Cited By ~ 16

Author(s):

Q Yang ◽

L Hanson ◽

M D Bennett ◽

I J Leitch

Keyword(s):

Ribosomal Dna ◽

Genomic Structure ◽

In Situ Hybridisation ◽

Avena Fatua ◽

The Other ◽

Morphological Data ◽

Avena Species ◽

Intergenomic Translocations ◽

Genomic In Situ Hybridisation

Allohexaploid wild oat, Avena fatua L. (Poaceae; 2n = 6x = 42), is one of the world's worst weeds, yet unlike some of the other Avena hexaploids, its genomic structure has been relatively little researched. Consequently, in situ hybridisation was carried out on one accession of A. fatua using an 18S-25S ribosomal DNA (rDNA) sequence and genomic DNA fromA. strigosa (AA-genome diploid) and A. clauda (CC-genome diploid) as probes. Comparing these results with those for other hexaploids studied previously: (i) confirmed that the genomic composition of A. fatua was similar to the other hexaploid Avena taxa (i.e., AACCDD), (ii) identified major sites of rDNA on three pairs of A/D-genome chromosomes, in common with other Avena hexaploids, and (iii) revealed eight chromosome pairs carrying intergenomic translocations between the A/D- and C-genomes in the accession studied. Based on karyotype structure, the identity of some of these recombinant chromosomes was proposed, and this showed that some of these could be divided into two types, (i) those common to all hexaploid Avena species analysed (3 translocations) and (ii) one translocation in this A. fatua accession not previously observed in reports on other hexaploid Avena species. If this translocation is found to be unique to A. fatua, then this information, combined with more traditional morphological data, will add support to the view that A. fatua is genetically distinct from other hexaploid Avena species and thus should retain its full specific status.Key words: wild oats, Avena, genomic in situ hybridisation (GISH), intergenomic translocations, ribosomal DNA.

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A COMPARISON OF CHROMOSOME NUMBER AND KARYOTYPE IN SOMATIC CHROMOSOMES OF STANGERIACEAE (CYCADALES)

Edinburgh Journal of Botany ◽

10.1017/s0960428601000786 ◽

2001 ◽

Vol 58 (3) ◽

pp. 475-481 ◽

Cited By ~ 6

Author(s):

G. KOKUBUGATA ◽

K. D. HILL ◽

G. W. WILSON ◽

K. KONDO ◽

L. M. RANDALL

Keyword(s):

Chromosome Number ◽

Staining Method ◽

Karyotype Analysis ◽

Mitotic Metaphase ◽

Chromosomal Change ◽

First Time ◽

Orcein Staining

Somatic chromosomes at mitotic metaphase of two species and two undescribed populations of Bowenia, and Stangeria eriopus, which were classified in Stangeriaceae, Cycadales, were compared using the standard aceto-orcein staining method. All Bowenia taxa showed a chromosome number of 2n = 18, while S. eriopus showed a chromosome number of 2n = 16. The chromosome number of 2n = 18 in B. ‘Kuranda’ is reported for the first time. The present karyotype analysis indicates that B. ‘Kuranda’ and another undescribed taxon, B. ‘Tinaroo’, are cytotaxonomically closer to B. spectabilis than B. serrulata, and that the karyotype of Stangeria is unlikely to have been derived from that of Bowenia by a simple chromosomal change such as centromeric fission and deletion.

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Chromosome in situ hybridisation of ribosomal DNA in Erianthus sect. Ripidium species with varying chromosome numbers confirms x = 10 in Erianthus sect. Ripidium

Genome ◽

10.1139/g98-126 ◽

1999 ◽

Vol 42 (2) ◽

pp. 270-273 ◽

Cited By ~ 2

Author(s):

P Besse ◽

C L McIntyre

Keyword(s):

Chromosome Number ◽

Ribosomal Dna ◽

Chromosome Numbers ◽

In Situ Hybridisation ◽

Basic Chromosome Number ◽

Dna Probe ◽

Published Data ◽

Basic Chromosome ◽

Fluorescent In Situ Hybridisation

A wheat ribosomal DNA probe was used to determine the number of rDNA-carrying chromosomes in 2 Erianthus sect. Ripidium species using FISH (fluorescent in situ hybridisation) and non-fluorescent ISH. Two and four ribosomal DNA sites were revealed in E. elephantinus (2n = 20) and E. procerus (2n = 40), respectively. This result, together with previously published data showing 6 rDNA-carrying chromosomes in E. arundinaceus (2n = 60), confirms a possible basic chromosome number of x = 10 in Erianthus sect. Ripidium.Key words: Erianthus, FISH, ISH, ribosomal DNA, Saccharum, sugarcane.

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Genome size, chromosome number, and rDNA organisation in Algerian populations of Artemisia herba-alba (Asteraceae), a basic plant for animal feeding facing overgrazing erosion

Anales del Jardín Botánico de Madrid ◽

10.3989/ajbm.2415 ◽

2016 ◽

Vol 73 (2) ◽

pp. 043

Author(s):

Youcef Bougoutaia ◽

Sònia Garcia ◽

Teresa Garnatje ◽

Meriem Kaid-Harche ◽

Joan Vallès

Keyword(s):

Chromosome Number ◽

Genome Size ◽

In Situ Hybridisation ◽

Fluorescence In Situ Hybridisation ◽

Ploidy Levels ◽

Rdna Genes ◽

Basic Plant ◽

Tetraploid Cytotypes ◽

First Time

Artemisia herba-alba is a largely-distributed and often landscape-dominating taxon in arid areas of the Mediterranean and Irano-Turanian regions. In Algeria, in 2010 its communities covered 10% of the steppe territory, but its populations have been subjected to overgrazing. A karyological study based on 22 populations together with a cytogenetic characterisation of this species has been performed for the first time in Algerian materials, through genome size and chromosome number determination. Fluorescence in situ hybridisation (FISH) was also used to assess the rDNA loci number and distribution in the two ploidy levels detected. The studied accessions are diploid (2n = 2x = 18 chromosomes, 6 populations) or tetraploid (2n = 4x = 36 chromosomes, 15 populations). One population, occupying a more or less central geographic position among the studied area, presented both cytotypes. Genome size reflects well the two ploidy levels, with no evidence of downsizing with polyploidy. The karyotypes are rather symmetric (2A Stebbins’ class). FISH analyses detected four signals (2 loci) in diploid and eight signals (4 loci) in tetraploid cytotypes for both ribosomal DNA genes, which present an L-type (linked) organisation, i.e. with loci from both rDNA genes colocalised. The presence of two ploidy levels suggest a genomic dynamism and even a possible differentiation underlying the morphological uniformity and despite the dramatic decrease experienced by this plant in Algeria in terms of surface coverage.

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Cytogenetical and cytotaxonomical analysis of some Brazilian species of Eleocharis (Cyperaceae)

Australian Journal of Botany ◽

10.1071/bt07017 ◽

2008 ◽

Vol 56 (1) ◽

pp. 82 ◽

Cited By ~ 10

Author(s):

Carlos Roberto Maximiano da Silva ◽

Maria Socorro González-Elizondo ◽

Letícia do Nascimento Andrade de Almeida Rego ◽

José Marcelo Domingues Torezan ◽

André Luís Laforga Vanzela

Keyword(s):

Karyotype Analysis ◽

In Situ Hybridisation ◽

Basic Number ◽

Fluorescence In Situ Hybridisation ◽

Chromosome Size ◽

45S Rdna ◽

Rdna Probe ◽

Parallel Movement ◽

Frequent Event

Karyotype analysis of 21 samples of 11 species of Eleocharis (Cyperaceae) from 10 localities in Brazil, showed the presence of chromosomes without primary constrictions and parallel movement of chromatids at metaphase–anaphase transition. Only the terminal nucleolar constrictions (satellites) were visualised. The chromosome numbers varied from 2n = 6 in E. subarticulata to 2n = 54 in E. acutangula, but the chromosome basic number x = 5 was confirmed. Generally, C-CMA3+ bands appear mostly in the extremities of the chromosomes, associated to NOR, and interstitial C-CMA3 bands were found only in E. geniculata and E. acutangula. C-DAPI+ bands were not found. Fluorescence in situ hybridisation (FISH) with the 45S rDNA probe was performed in five species. The results showed from four to eight hybridisation signals, always terminal. The analysed species include representatives of the following three subgenera of Eleocharis that occur in Brazil: Limnochloa, Scirpidium and Eleocharis. Species from the subgenus Limnochloa have small and numerous chromosomes. The remaining species, belonging to subgenera Eleocharis and Scirpidium, possess fewer and larger chromosomes. In subgenus Eleocharis, karyotypes of the section Eleocharis were differentiated by symploidy, agmatoploidy and polyploidy, whereas species of the section Eleogenus were all polyploids. Polyploidy seems to be the most frequent event in the karyotype differentiation in Eleocharis, but changes in the chromosome size and repetitive DNA sites were also observed.

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Human papilloma virus detection by in situ hybridisation signal amplification based on biotinylated tyramine deposition

Molecular Pathology ◽

10.1136/mp.49.6.m340 ◽

1996 ◽

Vol 49 (6) ◽

pp. M340-M344 ◽

Cited By ~ 13

Author(s):

P J Poddighe ◽

J Bulten ◽

H M J Kerstens ◽

J C M Robben ◽

W J G Melchers ◽

...

Keyword(s):

Human Papilloma Virus ◽

Signal Amplification ◽

In Situ Hybridisation ◽

Virus Detection ◽

Papilloma Virus ◽

Hybridisation Signal

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Bayesian fluorescence in situ hybridisation signal classification

Artificial Intelligence in Medicine ◽

10.1016/j.artmed.2003.11.005 ◽

2004 ◽

Vol 30 (3) ◽

pp. 301-316 ◽

Cited By ~ 25

Author(s):

Boaz Lerner

Keyword(s):

In Situ Hybridisation ◽

Signal Classification ◽

Fluorescence In Situ Hybridisation ◽

Hybridisation Signal

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Quantum Dot Based Duplex In Situ Hybridisation for Gene Expression Profiling.

Blood ◽

10.1182/blood.v106.11.3265.3265 ◽

2005 ◽

Vol 106 (11) ◽

pp. 3265-3265

Author(s):

Eleni Tholouli ◽

Dolores Di Vizio ◽

Fionnuala O’Connell ◽

Massimo Loda ◽

David Twomey ◽

...

Keyword(s):

Bone Marrow ◽

Myeloid Leukaemia ◽

Semiconductor Nanocrystals ◽

In Situ Hybridisation ◽

Spectral Imaging ◽

Quantitative Detection ◽

Oligonucleotide Probes ◽

Bone Marrow Trephine ◽

Hybridisation Signal

Abstract Quantum dots (QDs) are fluorescent semiconductor nanocrystals (2–10-nm core diameter) possessing the unique properties of extremely high fluorescence efficiency, lack of photobleaching and long fluorescence lifetime, making them an ideal tool for bioimaging. We have developed a novel technique for in situ hybridisation (ISH) using biotinylated oligonucleotides conjugated to streptavidin coated QD, and used them in this study to label bone marrow trephine samples. 50-mer long oligonucleotide probes were conjugated to QDs prior to ISH and conjugation efficiency was demonstrated by gel electrophopresis. ISH conditions and molar ratio of QDs to probe were optimised using a polyT probe. Images were captured using a CRI Nuance spectral imaging system and signal intensity was semi-quantitated using IPLab software. Specific oligonucleotide hybridisation was demonstrated using a probe for myeloperoxidase (MPO) in 40 bone marrow sections infiltrated by acute myeloid leukaemia (AML), acute lymphoblastic leukaemia (ALL), chronic myeloid leukaemia (CML) as well as reactive bone marrow. In each case hybridisation signal was consistent with the distribution of MPO by standard immunohistochemistry - MPO was strongly expressed by myeloid blasts and absent in lymphoid blasts; in CML, most, but not all, cells were positive for MPO, in comparison to many fewer positive cells in reactive marrow. Duplex ISH was demonstrated using a probe for bcl-2 together with MPO in 5 bone marrow sections infiltrated by follicular lymphoma (FL). Strong hybridisation signal for bcl-2 was detected in all cells of the paratrabecular aggregates of FL but showed only scattered positivity in the remainder of the bone marrow. Conversely, MPO was absent in the paratrabecular aggregates and present in the myeloid cells in the remainder of the marrow. This pattern was present in both single and duplex ISH for MPO and bcl-2 in the marrow infiltrated by FL. Duplex ISH was performed both by sequential hybridisation with bcl-2 followed by MPO, and simultaneously with a mix of bcl-2 and MPO probes. As negative controls, scrambled oligonucleotide probes for the corresponding genes were used in each case and did not show hybridisation. In summary, we have developed a generic method for QD labelling and semi-quantitative detection of oligonucleotide ISH in routinely processed clinical tissue samples. Although, in this study we primarily used bone marrow trephine samples, this technique can be applied to any tissue. It has the potential to facilitate transfer of microarray-identified gene signatures to clinical research and diagnostics, whilst the ability of spectral imaging to resolve multiple signals offers the possibility of multiplexed probe detection.

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