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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 2n = 2x = 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

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                gmb
                Genetics and Molecular Biology
                Genet. Mol. Biol.
                Sociedade Brasileira de Genética (Ribeirão Preto )
                1678-4685
                2009
                : 32
                : 2
                : 320-327
                Affiliations
                [1 ] Universidade Estadual de Londrina Brazil
                Article
                S1415-47572009000200019
                10.1590/S1415-47572009000200019
                4ed9a488-7fca-4935-a13f-12408d0a5aad

                http://creativecommons.org/licenses/by/4.0/

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                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=1415-4757&lng=en
                Categories
                BIOCHEMISTRY & MOLECULAR BIOLOGY
                GENETICS & HEREDITY

                Molecular biology,Genetics
                Cestrum,chromosome banding,FISH,heterochromatin,physical maps,45S and 5S rDNA
                Molecular biology, Genetics
                Cestrum, chromosome banding, FISH, heterochromatin, physical maps, 45S and 5S rDNA

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