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  • Fluorescence in situ hybridization  (2)
  • Alliaceae  (1)
  • Key words Chromosome elongation  (1)
  • Springer  (4)
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  • Springer  (4)
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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 93 (1996), S. 477-484 
    ISSN: 1432-2242
    Keywords: Key words Microdissection of plant centromeres ; Subtractive hybridization ; Fluorescence in situ hybridization ; Indirect immunofluorescence ; Immunoadsorption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The centromeric region of a telocentric field bean chromosome that resulted from centric fission of the metacentric satellite chromosome was microdissected. The DNA of this region was amplified and biotinylated by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR)/linker-adapter PCR. After fluorescence in situ hybridization (FISH) the entire chromosome complement of Vicia faba was labelled by these probes except for the nucleolus organizing region (NOR) and the interstitial heterochromatin, the chromosomes of V. sativa and V. narbonensis were only slightly labelled by the same probes. Dense uniform labelling was also observed when a probe amplified from a clearly delimited microdissected centromeric region of a mutant of Tradescantia paludosa was hybridized to T. paludosa chromosomes. Even after six cycles of subtractive hybridization between DNA fragments amplified from centromeric and acentric regions no sequences specifically located at the field bean centromeres were found among the remaining DNA. A mouse antiserum was produced which detected nuclear proteins of 33 kDa and 68 kDa; these were predominantly located at V. faba kinetochores during mitotic metaphase. DNA amplified from the chromatin fraction adsorbed by this serum out of the sonicated total mitotic chromatin also did not cause specific labelling of primary constrictions. From these results we conclude: (1) either centromere-specific DNA sequences are not very conserved among higher plants and are – at least in species with large genomes – intermingled with complex dispersed repetitive sequences that prevent the purification of the former, or (2) (some of) the dispersed repeats themselves specify the primary constrictions by stereophysical parameters rather than by their base sequence.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 96 (1998), S. 1022-1026 
    ISSN: 1432-2242
    Keywords: Key words Chromosome elongation ; Deletion ; Mitosis ; Disturbed development ; Fertility ; Apoptosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Chromosomes elongated beyond a critical size by balanced rearrangements reduce the viability and fertility of field bean individuals. The severity of symptoms, ranging from growth retardation to early death of seedlings, increases with the length of the longest chromosome arm. This is paralleled by the incomplete separation of sister chromatids during nuclear division, resulting in chromatin connections between daughter nuclei which become disrupted by cell-wall formation and yield chromatid deletions detectable as micronuclei. By means of the TUNEL assay we show that, compared to the wild-type, a 〉6-fold higher number of meristematic cells of karyotypes with chromosome arms surpassing the limit of tolerance reveal apoptotic nuclei and are prone to die. Thus, terminal chromatid deletions apparently trigger unscheduled apoptosis. Extensive cell death in meristems is eventually responsible for reduced growth, disturbed development and reduced seed set. Differentiated root tissues and microspores did not reveal apoptotic nuclei.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 93 (1996), S. 477-484 
    ISSN: 1432-2242
    Keywords: Microdissection of plant centromeres ; Subtractive hybridization ; Fluorescence in situ hybridization ; Indirect immunofluorescence ; Immunoadsorption
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The centromeric region of a telocentric field bean chromosome that resulted from centric fission of the metacentric satellite chromosome was microdissected. The DNA of this region was amplified and biotinylated by degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR)/linker-adapter PCR. After fluorescence in situ hybridization (FISH) the entire chromosome complement of Vicia faba was labelled by these probes except for the nucleolus organizing region (NOR) and the interstitial heterochromatin, the chromosomes of V. sativa and V. narbonensis were only slightly labelled by the same probes. Dense uniform labelling was also observed when a probe amplified from a clearly delimited microdissected centromeric region of a mutant of Tradescantia paludosa was hybridized to T. paludosa chromosomes. Even after six cycles of subtractive hybridization between DNA fragments amplified from centromeric and acentric regions no sequences specifically located at the field bean centromeres were found among the remaining DNA. A mouse antiserum was produced which detected nuclear proteins of 33 kDa and 68 kDa; these were predominantly located at V. faba kinetochores during mitotic metaphase. DNA amplified from the chromatin fraction adsorbed by this serum out of the sonicated total mitotic chromatin also did not cause specific labelling of primary constrictions. From these results we conclude: (1) either centromere-specific DNA sequences are not very conserved among higher plants and are — at least in species with large genomes — intermingled with complex dispersed repetitive sequences that prevent the purification of the former, or (2) (some of) the dispersed repeats themselves specify the primary constrictions by stereophysical parameters rather than by their base sequence.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Plant systematics and evolution 202 (1996), S. 255-264 
    ISSN: 1615-6110
    Keywords: Alliaceae ; Allium ; Satellite DNA ; fluorescent in situ hybridization ; sequence homology ; phylogeny
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A satellite sequence repeat ofAllium cepa was tested by fluorescent in situ hybridization (FISH) for cross-hybridization to chromosomes of 27 species (in 37 accessions) belonging to 14 sections of four subgenera ofAllium. All investigated species of sect.Cepa, with the two subsects.Cepa andPhyllodolon, revealed clear satellite-specific hybridization signals mainly at their chromosome termini. The tested species belonging to other sections/subgenera revealed no hybridization signals. An exception wasA. roylei, assigned to sect.Oreiprason. Its chromosomes also showed strong terminal hybridization signals. This and other features suggest a close relationship ofA. roylei to the species of sect.Cepa in spite of deviating morphological characters. The divergence between the satellite repeats to species to which theA. cepa repeat cross-hybridized was determined and revealed high degrees of similarity. Therefore, we conclude that this satellite sequence had evolved already in progenitor forms of sect.Cepa and remained unusually well conserved during speciation. This might indicate selection pressure exerted on a secondarily acquired telomere function of the satellite sequence.
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