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      Karyotype differentiation of four Cestrum species (Solanaceae) revealed by fluorescent chromosome banding and FISH

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          Abstract

          The karyotypes of four South American species of Cestrum ( C. capsulare, C. corymbosum, C. laevigatum and C. megalophylum) were studied using conventional staining, C-CMA/DAPI chromosome banding and FISH with 45S and 5S rDNA probes. The karyotypes showed a chromosome number of 2 n = 2 x = 16, with metacentric chromosomes, except for the eighth submeta- to acrocentric pair. Several types of heterochromatin were detected, which varied in size, number, distribution and base composition. The C-CMA + bands and 45S rDNA were located predominantly in terminal regions. The C-CMA + /DAPI + bands appeared in interstitial and terminal regions, and the C-DAPI + bands were found in all chromosome regions. The 5S rDNA sites were observed on the long arm of pair 8 in all species except C. capsulare, where they were found in the paracentromeric region of the long arm of pair 4. The differences in band patterns among the species studied here, along with data from other nine species reported in the literature, suggest that the bands are dispersed in an equilocal and non-equilocal manner and that structural rearrangements can be responsible for internal karyotype diversification. However, it is important to point out that the structural changes involving repetitive segments did not culminate in substantial changes in the general karyotype structure concerning chromosome size and morphology.

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          Cloning and characterization of ribosomal RNA genes from wheat and barley.

          Wheat and barley DNA enriched for ribosomal RNA genes was isolated from actinomycin D-CsCl gradients and used to clone the ribosomal repeating units in the plasmid pAC184. All five chimeric plasmids isolated which contained wheat rDNA and eleven of the thirteen which had barley rDNA were stable and included full length ribosomal repeating units. Physical maps of all length variants cloned have been constructed using the restriction endonucleases Eco Rl, Bam Hl, Bgl II, Hind III and Sal I. Length variation in the repeat units was attributed to differences in the spacer regions. Comparison of Hae III and Hpa II digestion of cereal rDNAs and the cloned repeats suggests that most methylated cytosines in natural rDNA are in -CpG-. Incomplete methylation occurs at specific Bam Hl sites in barley DNA. Detectable quantities of ribosomal spacer sequences are not present at any genomic locations other than those of the ribosomal RNA gene repeats.
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            Sequence organization of the repeating units in the nucleus of wheat which contain 5S rRNA genes.

            Examples of both the 410 and 500 bp size classes of repeating units containing wheat 5S rRNA genes have been cloned in plasmid pBR322 and sequenced. The structural genes showed sequence microheterogeneity. Also the gene in the 500 bp repeat which was sequenced had a 15 bp tandem duplication within it and appears to be representative of non-transcribed subfamily of repeating units. The transcription terminators comprise 14-17 A.T bp immediately preceded by a region of weak dyad symmetry. The spacer regions adjacent to the transcription terminators in the two different size repeat units have interspersed oligonucleotides of high and low homology. The central spacer regions of the two size classes have very different sequences. The only repeated sequence in the spacers has undergone extensive divergence. In contrast to most of the spacer, the 70 bp region preceding the genes of each type of repeat show high homology, suggesting that it has functional importance. The transcription start point obeys the pyrimidine-1 purine+1 rule.
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              Integrated cytogenetic map of chromosome arm 4S of A. thaliana: structural organization of heterochromatic knob and centromere region.

              We have constructed an integrated cytogenetic map of chromosome arm 4S of Arabidopsis thaliana. The map shows the detailed positions of various multicopy and unique sequences relative to euchromatin and heterochromatin segments. A quantitative analysis of the map positions at subsequent meiotic stages revealed a striking pattern of spatial and temporal variation in chromatin condensation for euchromatin and heterochromatin. For example, the centromere region consists of three domains with distinguishable structural, molecular, and functional properties. We also characterized a conspicuous heterochromatic knob of approximately 700 kb that accommodates a tandem repeat and several dispersed pericentromere-specific repeats. Moreover, our data provide evidence for an inversion event that relocated pericentromeric sequences to an interstitial position, resulting in the heterochromatic knob.
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                Author and article information

                Journal
                Genet Mol Biol
                GMB
                Genetics and Molecular Biology
                Sociedade Brasileira de Genética (Ribeirão Preto, SP, Brazil )
                1415-4757
                1678-4685
                Apr-Jun 2009
                1 June 2009
                : 32
                : 2
                : 320-327
                Affiliations
                [1]simpleLaboratório de Biodiversidade e Restauração de Ecossistemas, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina, PR Brazil
                Author notes
                Send correspondence to André Luís Laforga Vanzela. Laboratório de Biodiversidade e Restauração de Ecossistemas, Centro de Ciências Biológicas, Universidade Estadual de Londrina, Caixa Postal 6001, 86051-990 Londrina, PR, Brazil. E-mail: andrevanzela@ 123456uel.br .
                Article
                10.1590/S1415-47572009000200019
                3036934
                21637687
                4ed9a488-7fca-4935-a13f-12408d0a5aad
                Copyright © 2009, Sociedade Brasileira de Genética.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 2 May 2008
                : 8 September 2008
                Categories
                Plant Genetics
                Research Article

                Molecular biology
                physical maps,heterochromatin,45s and 5s rdna,chromosome banding, cestrum,fish
                Molecular biology
                physical maps, heterochromatin, 45s and 5s rdna, chromosome banding, cestrum, fish

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