Dynamic Changes in the Distribution of a Satellite Homologous to Intergenic 26-18S rDNA Spacer in the Evolution of Nicotiana

Genetics ◽  
2004 ◽  
Vol 166 (4) ◽  
pp. 1935-1946
Author(s):  
K Y Lim ◽  
K Skalicka ◽  
B Koukalova ◽  
R A Volkov ◽  
R Matyasek ◽  
...  

Abstract An ∼135-bp sequence called the A1/A2 repeat was isolated from the transcribed region of the 26-18S rDNA intergenic spacer (IGS) of Nicotiana tomentosiformis. Fluorescence in situ hybridization (FISH) and Southern blot analysis revealed its occurrence as an independent satellite (termed an A1/A2 satellite) outside of rDNA loci in species of Nicotiana section Tomentosae. The chromosomal location, patterns of genomic dispersion, and copy numbers of its tandemly arranged units varied between the species. In more distantly related Nicotiana species the A1/A2 repeats were found only at the nucleolar organizer regions (NOR). There was a trend toward the elimination of the A1/A2 satellite in N. tabacum (tobacco), an allotetraploid with parents closely related to the diploids N. sylvestris and N. tomentosiformis. This process may have already commenced in an S3 generation of synthetic tobacco. Cytosine residues in the IGS were significantly hypomethylated compared with the A1/A2 satellite. There was no clear separation between the IGS and satellite fractions in sequence analysis of individual clones and we found no evidence for CG suppression. Taken together the data indicate a dynamic nature of the A1/A2 repeats in Nicotiana genomes, with evidence for recurrent integration, copy number expansions, and contractions.

2019 ◽  
Vol 74 (11-12) ◽  
pp. 319-328
Author(s):  
Oleg Georgiev ◽  
Kiril Mishev ◽  
Maria Krasnikova ◽  
Meglena Kitanova ◽  
Anna Dimitrova ◽  
...  

Abstract Hordeum vulgare and Hordeum bulbosum are two closely related barley species, which share a common H genome. H. vulgare has two nucleolar organizer regions (NORs), while the NOR of H. bulbosum is only one. We sequenced the 2.5 kb 25S-18S region in the rDNA of H. bulbosum and compared it to the same region in H. vulgare as well as to the other Triticeae. The region includes an intergenic spacer (IGS) with a number of subrepeats, a promoter, and an external transcribed spacer (5′ETS). The IGS of H. bulbosum downstream of 25S rRNA contains two 143-bp repeats and six 128-bp repeats. In contrast, the IGS in H. vulgare contains an array of seven 79-bp repeats and a varying number of 135-bp repeats. The 135-bp repeats in H. vulgare and the 128-bp repeats in H. bulbosum show similarity. Compared to H. vulgare, the 5′ETS of H. bulbosum is shorter. Additionally, the 5′ETS regions in H. bulbosum and H. vulgare diverged faster than in other Triticeae genera. Alignment of the Triticeae promoter sequences suggests that in Hordeum, as in diploid Triticum, transcription starts with guanine and not with adenine as it is in many other plants.


Genome ◽  
1996 ◽  
Vol 39 (2) ◽  
pp. 410-417 ◽  
Author(s):  
A. Katsiotis ◽  
T. Schmidt ◽  
J. S. Heslop-Harrison

A cloned repetitive sequence, pAvKB30, obtained from an Avena vaviloviana (AB genome) genomic library, along with two polymerase chain reaction products derived from the conserved region of the reverse transcriptase (RT) gene of retrotransposons, were characterized molecularly and cytologically. The cloned DNA fragment was a dispersed repeat present in all Avena species used in this study (A. strigosa, A. clauda, A. vaviloviana, A. magna, and A. sativa). The fragment was sequenced (210 bp) and found to be 69.5% homologous to part of WIS-2-1A, and 60.5% homologous to the leader sequence of BARE-1; both of these elements have been characterized as Ty1-copia-like retrotransposons in wheat and barley, respectively. In situ hybridization of pAvKB30 to diploid, tetraploid, and hexaploid oat species revealed that the probe is present on both arms of all chromosomes (A, B, C, and D genomes) but is excluded from their centromeric and nucleolar organizer regions. By using double in situ hybridization in hexaploid A. sativa (ACD genome), pAvKB30 was found to be present in lower copy numbers in C-genome chromosomes compared with A- and D-genome chromosomes. Furthermore, under low stringency conditions, pAvKB30 hybridized on Southern blots containing barley, wheat, rye, and Arrhenatherum DNA. However, under high stringency conditions, it hybridized only on Arrhenatherum DNA, which is considered to be the genus most closely related to Avena. All Avena species included in this study yielded a PCR product when the primers from the RT domain of retrotransposons were used. Two products, rtA, obtained by using A. strigosa (As genome) as template, and rtC, obtained by using A. clauda (Cp genome) as template, gave Southern and in situ hybridization results similar to pAvKB30, but each was more abundant in its genome of origin. Key words : genomes, oats, in situ hybridization, translocations, repetitive sequence.


1998 ◽  
Vol 111 (10) ◽  
pp. 1433-1439
Author(s):  
F. Zurita ◽  
R. Jimenez ◽  
M. Burgos ◽  
R.D. de la Guardia

We have developed a procedure for sequential silver staining and in situ hybridization to analyze the relationship between the amount of rDNA present in nucleolar organizer regions, as estimated by in situ hybridization, and their level of expression, as estimated by the silver signal. For simplicity we used cells from the insectivorous mole Talpa occidentalis, which have a single pair of nucleolar organizer regions in chromosome pair 3. The relative content of ribosomal cistrons was also related to the hierarchy of activation of the nucleolar organizer regions present in this chromosomal pair. Statistical analyses demonstrated that both the relative level of expression and the activation hierarchy depended mainly on the number of ribosomal cistrons in nucleolar organizer regions. We propose a functional two-step hypothesis, which is consistent with most known data concerning interchromosomal, intercellular and interindividual variation in a number of plant and animal species, including Talpa occidentalis. In step one, the first available transcription factors bind randomly to the ribosomal promoters, such that larger nucleolar organizer regions are more likely to recruit them. In the second step the remaining transcription factors are recruited in a cooperative way, thus completing activation of one nucleolar organizer region, before the next one becomes active.


2000 ◽  
Vol 20 (4) ◽  
pp. 155-162 ◽  
Author(s):  
Lukas Günther ◽  
Peter Hufnagl ◽  
Klaus‐Jürgen Winzer ◽  
Hans Guski

The relation between estrogen receptors (ER) and argyrophilic nucleolar organizer regions (AgNORs)in situwithin human breast cancer cells was analyzed. For AgNOR measurements in 49 invasive breast carcinomas, a new reproducible staining method for dual demonstration of ER and AgNORs was applied. Quantitative AgNOR variables were determined in ER‐positive and ER‐negative tumor cells by digital image analysis. The relationships between AgNOR parameters of ER‐positive and ER‐negative cells and other prognostic factors of breast cancer [Bloom–Richardson‐Grading and growth fraction (Ki‐67 index)] were investigated. A higher AgNOR content in ER‐negative cells and a special clustering phenomenon in ER‐positive tumor cells were found. Correlation with other criteria of malignant potential could be exclusively demonstrated for ER‐negative cells. ER‐negative cells of breast cancer can be characterized as the more malignant and possibly prognosis‐dictating cell fraction. Thus, ER‐negative cells probably contribute more to the progression of the tumor disease and furthermore to the prognosis than ER‐positive cells. We recommend measurement AgNORs exclusively in ER‐negative cells of breast cancer.


Genome ◽  
2010 ◽  
Vol 53 (2) ◽  
pp. 125-137 ◽  
Author(s):  
Ekaterina D. Badaeva ◽  
Olga Yu. Shelukhina ◽  
Axel Diederichsen ◽  
Igor G. Loskutov ◽  
Vitaly A. Pukhalskiy

The chromosome set of Avena macrostachya Balansa ex Coss. et Durieu was analyzed using C-banding and fluorescence in situ hybridization with 5S and 18S-5.8S-26S rRNA gene probes, and the results were compared with the C-genome diploid Avena L. species. The location of major nucleolar organizer regions and 5S rDNA sites on different chromosomes confirmed the affiliation of A. macrostachya with the C-genome group. However, the symmetric karyotype, the absence of “diffuse heterochromatin”, and the location of large C-band complexes in proximal chromosome regions pointed to an isolated position of A. macrostachya from other Avena species. Based on the distribution of rDNA loci on the C-genome chromosomes of diploid and polyploid Avena species, we propose a model of the chromosome alterations that occurred during the evolution of oat species.


PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5316 ◽  
Author(s):  
Naiara Pereira Araújo ◽  
Cayo Augusto Rocha Dias ◽  
Rodolfo Stumpp ◽  
Marta Svartman

Trinomys Thomas (1921) is a terrestrial genus of spiny rats endemic to the Brazilian areas of Atlantic Forest and the transitional areas of Cerrado and Caatinga. Although most species have been already karyotyped, the available cytogenetic information is mostly restricted to diploid and fundamental numbers. We analyzed the chromosomes of two Trinomys species: Trinomys moojeni (2n = 56, FN = 106) and Trinomys setosus setosus (2n = 56, FN = 106 and 2n = 56, FN = 108). Our analyses included GTG- and CBG-banding, silver-staining of the nucleolar organizer regions, and chromosome mapping of telomeres and 45S rDNA by fluorescent in situ hybridization (FISH). Comparative GTG- and CBG-banding suggested that the interspecific variation may be due to rearrangements such as pericentric inversions, centromere repositioning, and heterochromatin variation. We report two new karyotypes for T. s. setosus and describe for the first time the banding patterns of the two Trinomys species.


2006 ◽  
Vol 66 (1b) ◽  
pp. 191-198 ◽  
Author(s):  
R. F. Artoni ◽  
M. L. Terêncio ◽  
M. R. Vicari ◽  
M. C. A. Matiello ◽  
M. M. Cestari ◽  
...  

Karyotypic data are presented for two sympatric Corydoras species of the Lagoa Dourada, namely, C. ehrhadti and C. paleatus, which are found in the upper Tibagi river basin (Ponta Grossa, State of Paraná, Brazil). The same diploid number and karyotypic formula were observed in both species/populations. A great similarity in the constitutive heterochromatin distribution and in the activity of nucleolar organizer regions was also found. The use of in situ hybridization with a fluorescent 18S rDNA probe allowed for the identification of the species/populations through the location of ribosomal sites.


2007 ◽  
Vol 67 (2) ◽  
pp. 333-338 ◽  
Author(s):  
LVS De Rosa ◽  
F. Foresti ◽  
C. Martins ◽  
C. Oliveira ◽  
PE. Sobrinho ◽  
...  

Cytogenetic analyses were performed in two Curimatidae species (Steindachnerina insculpta and Cyphocharax modesta) from the Paranapanema and Tietê Rivers (São Paulo State, Brazil), showing a karyotype composed of 54 meta-submetacentric chromosomes in both species. Silver- and chromomycyn-staining and fluorescent in situ hybridization (FISH) using a 18S rDNA probe indicated that the nucleolar organizer regions (NORs) of both species are localized in the terminal region of the long arm of two metacentric chromosomes. Although a single NOR system was evidenced in both analyzed species, S. insculpta and C. modesta presented the nucleolar organizer regions in distinct chromosome pairs, indicating that these cistrons can be considered cytogenetic markers. Variation on the amount and distribution of the constitutive heterochromatin (C-bands) could also be detected between the two species - while S. insculpta presented few heterochromatic blocks, intensely stained C-bands were evidenced in C. modesta specially in the terminal region of the long arm of the NOR-bearing chromosomes. Although most Curimatidae species have been characterized by homogeneous karyotypes, isolated populations could be established under different environmental conditions leading to karyotype micro-structure variations specially related to the NORs localization and C-banding distribution. The obtained data were useful for the cytogenetic characterization and differentiation of S. insculpta and C. modesta and could be used in evolutionary inferences in the Curimatidae group.


2009 ◽  
Vol 52 (4) ◽  
pp. 893-899
Author(s):  
Aline Dias Brandão ◽  
Lyderson Facio Viccini ◽  
Shirlei Maria Recco-Pimentel

Since previous cytogenetic reports of Aloysia have only described the meiotic behavior and chromosomal number of some species, the aim of this work was to provide detailed cytogenetic description of Aloysia virgata that would contribute to the understanding of the taxonomical organization of the Verbenaceae. Aloysia virgata had a karyotype with 2n = 36 metacentric chromosomes, all with similar size. The large amount of heterochromatin seen after Giemsa staining was confirmed by C-banding. Four nucleolar organizer regions (NORs) were detected with an rDNA 45S probe in two homologous pairs and two sites of 5S rDNA located on one chromosomal pair were detected by fluorescence in situ hybridization. The interphase nucleus was classified as semi-reticulate. Meiotic analysis showed a normal chromosomal behavior, with 18 bivalents in some parts of prophase I and in metaphase I. The number of chromosomes, NORs and 5S rDNA segments did not exclude a possible polyploid origin.


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