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      Characterization of the Arachis (Leguminosae) D genome using fluorescence in situ hybridization (FISH) chromosome markers and total genome DNA hybridization

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          Abstract

          Chromosome markers were developed for Arachis glandulifera using fluorescence in situ hybridization (FISH) of the 5S and 45S rRNA genes and heterochromatic 4'-6-diamidino-2-phenylindole (DAPI) positive bands. We used chromosome landmarks identified by these markers to construct the first Arachis species ideogram in which all the homologous chromosomes were precisely identified. The comparison of this ideogram with those published for other Arachis species revealed very poor homeologies with all A and B genome taxa, supporting the special genome constitution (D genome) of A. glandulifera. Genomic affinities were further investigated by dot blot hybridization of biotinylated A. glandulifera total DNA to DNA from several Arachis species, the results indicating that the D genome is positioned between the A and B genomes.

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          Most cited references40

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          Variation in satellite DNA profiles--causes and effects.

          Heterochromatic regions of the eukaryotic genome harbour DNA sequences that are repeated many times in tandem, collectively known as satellite DNAs. Different satellite sequences co-exist in the genome, thus forming a set called a satellite DNA library. Within a library, satellite DNAs represent independent evolutionary units. Their evolution can be explained as a result of change in two parameters: copy number and nucleotide sequence, both of them ruled by the same mechanisms of concerted evolution. Individual change in either of these two parameters as well as their simultaneous evolution can lead to the genesis of species-specific satellite profiles. In some cases, changes in satellite DNA profiles can be correlated with chromosomal evolution and could possibly influence the evolution of species.
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            The use of combined FISH/GISH in conjunction with DAPI counterstaining to identify chromosomes containing transgene inserts in amphidiploid tobacco.

            We have used combined fluorescent and genomic in situ hybridization (FISH/GISH) together with 4',6-diamidino-2-phenylindole (DAPI) counterstaining to determine simultaneously the chromosomal integration site and subgenomic allocation of a transgene insert in amphidiploid tobacco (Nicotiana tabacum, 2n = 4 chi = 48). The procedure provides sufficient information on physical markers to identify at least 20 out of 24 chromosome pairs of two tobacco cultivars commonly used in studies on transgene expression and silencing (cv. Petit Havana SR1 and cv. Gatersleben). The chromosomes can be distinguished on the basis of diploid parental ancestry, size, morphology, the presence of rDNA loci and/or intergenomic exchanges, and the DAPI banding pattern, which is shown here for the first time for N. tabacum. From a single ISH experiment, it should now be possible in most cases to identify a tobacco chromosome carrying a transgene insert, thus permitting systematic studies of how the chromosomal location of transgenes influences expression levels.
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              Use of WildArachisSpecies/Introgression of Genes intoA. hypogaea L.

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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Journal
                gmb
                Genetics and Molecular Biology
                Genet. Mol. Biol.
                Sociedade Brasileira de Genética (Ribeirão Preto )
                1678-4685
                2008
                : 31
                : 3
                : 717-724
                Affiliations
                [1 ] Instituto de Botánica del Nordeste Argentina
                [2 ] Universidad Nacional del Nordeste
                Article
                S1415-47572008000400019
                10.1590/S1415-47572008000400019
                e17d95ad-c48d-4b34-aa9d-4617e1aa97cd

                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
                chromosome markers,DAPI bands,rDNA loci,dot blot hybridization,genome relationships

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