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1

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Sequence organization and the mechanism of interstitial deletion clustering in a plant genome Vicia faba)

Schubert, I. ; Rieger, R. ; Fuchs, J. ; [et al.]

Amsterdam : Elsevier

Mutation Research Letters 325 (1994), S. 1-5

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ISSN: 0165-7992

Keywords: Aberration spectra ; Chromatid-type aberrations ; Deletion clustering ; Error-prone recombination repair ; Highly repetitive sequences ; Plant chromosomes

Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002

Topics: Biology , Medicine

Type of Medium: Electronic Resource

URL: http://linkinghub.elsevier.com/retrieve/pii/0165-7992(94)90020-5

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2

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Molecular-cytogenetic characterization of a higher plant centromere/kinetochore complex (1996)

Houben, A. ; Brandes, A. ; Pich, U. ; [et al.]

Springer

Theoretical and applied genetics 93 (1996), S. 477-484

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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.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/s001220050305

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3

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Unscheduled apoptosis in meristematic plant cells is triggered via terminal deletions in artificially elongated chromosome arms (1998)

Schubert, I. ; Oud, O. J. L. ; Pich, U.

Springer

Theoretical and applied genetics 96 (1998), S. 1022-1026

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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.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/s001220050834

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4

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Molecular-cytogenetic characterization of a higher plant centromere/kinetochore complex (1996)

Houben, A. ; Brandes, A. ; Pich, U. ; [et al.]

Springer

Theoretical and applied genetics 93 (1996), S. 477-484

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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.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00417938

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5

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Immunostaining and interphase arrangement of field bean kinetochores (1995)

Houben, A. ; Guttenbach, M. ; Kreß, W. ; [et al.]

Springer

Chromosome research 3 (1995), S. 27-31

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ISSN: 1573-6849

Keywords: CREST sera ; interphase kinetochore association ; kinetochore ; nuclear proteins ; Vicia faba

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Abstract More than 100 sera from patients with scleroderma CREST (calcinosis, Raynaud phenomenon, esophageal dismotility, sclerodactyly, telangiectasia) were tested in order to detect antigenic nuclear components of the field beanVicia faba (2n=12). Kinetochores of mitotic chromosomes and prekinetochores of interphase cells from root-tip meristems were specifically labelled via an indirect immunofluorescence procedure by antibodies of one of these sera. In 44% of interphase nuclei in which centromeres could be identified, only half (6) of the number of expected prekinetochores (12) was detected, circumstantially indicating at least transient association of homologous centromeres. Some nuclei showed clustering of centromeres at one pole (Rabl configuration). In metaphase chromosomes, each sister kinetochore contained a fluorescent spot. Western blotting of field bean nuclear proteins revealed four antigenic proteins of 28, 30, 64 and 68 kDa.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00711158

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6

Electronic Resource

Pich, U. ; Fritsch, R. ; Schubert, I.

Springer

Plant systematics and evolution 202 (1996), S. 255-264

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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.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00983386

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7

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Differentiation of field bean heterochromatin byin situ hybridization with a repeatedFokI sequence (1994)

Fuchs, J. ; Pich, U. ; Meister, A. ; [et al.]

Springer

Chromosome research 2 (1994), S. 25-28

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ISSN: 1573-6849

Keywords: chromosome banding ; FISH ; flow sorting ; FokI sequence repeats ; heterochromatin ; Vicia faba

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Abstract The chromosomes of a field bean line with a reconstructed karyotype (ACB) were hybridizedin situ with biotinylated probes of a repetitiveFok I sequence, of DOP-PCR (degenerate oligonucleotide primed polymerase chain reaction) amplified DNA from a chromosome that does not contain this sequence, and with probes containing dispersed repetitive sequences. The results were compared with Giemsa banding, DNA late replication andFokIin situ digestion patterns. This allowed further differentiation between the chromatin types of this species. Centromeric and NOR-associated heterochromatin as well as euchromatin were shown to be free ofFokI sequence repeats. Among the interstitial late replicating Giemsa bands, subdivided into ‘marker’ and ‘additional’ bands, most of the marker bands located at mid-arm positions were composed mainly or exclusively of tandemly arrangedFokI repeats. Some of the marker bands and nearly all of the additional bands located in the vicinity of centromeres were free ofFokI sequence repeats, ofFokI recognition sites, and possibly also of dispersed repetitive sequences. They are probably composed of specific, not yet defined, repetitive sequences.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF01539450

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