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  • 1
    Electronic Resource
    Electronic Resource
    Comparison of three DNA fluorochromes for flow cytometric estimation of nuclear DNA content in plants (1992)
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 85 (1992), S. 0 
    Details
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Flow cytometric estimation of nuclear DNA content was performed in six plant species employing three fluorochromes showing different DNA base preferences: propidium iodide (no base preference), 4′,6-diamidino-2-phenylindole (DAPI; AT preference), and mithramycin (GC preference). Nuclei isolated from human leukocytes were used as a primary reference standard. While nuclear DNA contents estimated using propidium iodide were in agreement with published data obtained using other techniques, the values obtained using fluorochromes showing base preference were significantly different. It was found that the differences were caused by the differences in overall AT/GC ratios, and by the species-specific differences in binding of these fluorochromes to DNA. It was concluded that nuclear DNA content estimations performed with fluorochromes showing base preference should be interpreted with caution even when AT/GC ratios of the reference and the sample are equal. The use of intercalting dyes (e.g. propidium iodide) is recommended for this purpose. On the other hand, comparison of the staining behaviour of intercalating dyes with that of dyes showing base preference may give additional information on chromatin structural differences and arrangement of molecule pairs in DNA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1399-3054.1992.tb04764.x
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  • 2
    Electronic Resource
    Electronic Resource
    Cytological study of interspecific hybrid between Brassica campestris x B. hirta (Sinapis alba) (1995)
    Springer
    Sexual plant reproduction 8 (1995), S. 37-43 
    Details
    ISSN: 1432-2145
    Keywords: Brassica hirta ; Brassica campestris ; Interspecific hybridization ; Ovary culture ; Flow cytometry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Interspecific hybrids from the crossing Brassica campestris x B. hirta are reported in our study for the first time. F1 plants were obtained by using ovary culture. The phenotype of hybrids was similar to B. napus; the plants were self-fertile. Investigation of meiotic division and nuclear DNA content measurements showed the amphidiploid origin of these hybrids. The relationship between genome A and D, as well as the spontaneous amphidiploidization of the hybrids, are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00228761
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  • 3
    Electronic Resource
    Electronic Resource
    Evolutionary conservation of kinetochore protein sequences in plants (2000)
    Springer
    Chromosoma 109 (2000), S. 482-489 
    Details
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The evolutionary conservation of structural/functional kinetochore proteins has been studied on isolated nuclei and pro-/metaphase chromosomes of mono- and dicot plants. The cross-reactivities of antibodies against human CENPC, CENPE and CENPF, and against maize CENPCa with the centromeric regions of mitotic chromosomes of Vicia faba and/or Hordeum vulgare are shown. Putative homologs of the kinetochore protein SKP1 (suppressor of kinetochore protein 1p of yeast) were found in both species and of CBF5p (centromere binding factor 5 of yeast) in barley. Antibodies against synthetic peptides derived from partial sequences encoding these proteins were produced and recognized the centromeric regions on mitotic chromosomes as detected by indirect immunofluorescence.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004120000109
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  • 4
    Electronic Resource
    Electronic Resource
    Localization of seed protein genes on flow-sorted field bean chromosomes (1993)
    Springer
    Chromosome research 1 (1993), S. 107-115 
    Details
    ISSN: 1573-6849
    Keywords: Vicia faba ; chromosome flow sorting ; gene mapping ; PCR ; seed protein genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chromosomes from reconstructed field bean (Vicia faba L.) karyotypes were flow-sorted and the DNA was used for the physical localization of seed storage and nonstorage (USP) protein genes using PCR with sequence specific primers. The data were confirmed and refined by using DNA of microisolated chromosomes of other karyotypes as the target for PCR. The specificity of the PCR products was proved by restrictase digestion into fragments of predicted length or by reamplification using ‘nested’ primers. The genes are located within defined regions of chromosome I (USP=unknown seedprotein genes), II (vicilin genes, legumin B3 genes), III (legumin B4 genes), IV (pseudogenes ψ1) and V (legumin A genes and pseudogenes ψ1). Except for the pseudogene derived from the sequence of legumin B4 gene, all members of each gene family are located in one chromosome region exclusively. This approach proved to be useful for localizing genes that cannot be mapped genetically (due to the lack of allelic variants) and might be applied to integrate physical and genetic maps.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00710033
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  • 5
    Electronic Resource
    Electronic Resource
    Relationship of embryogenic competence in maize (Zea mays L.) leaves to mitotic activity, cell cycle and nuclear DNA content (1992)
    Springer
    Plant cell, tissue and organ culture 31 (1992), S. 215-221 
    Details
    ISSN: 1573-5044
    Keywords: cell cycle ; DNA content ; embryogenic competence ; flow cytometry ; mitotic activity ; somatic embryogenesis ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Embryogenic competence of maize (Zea mays L.) leaf tissues was studied with regard to leaf age, distance from leaf base, mitotic activity, 2C nuclear DNA content, and the distribution of cells within the cell cycle. The highest embryogenic competence was observed in the first segment of the second innermost leaf. In a succession of 8 mm segments from the leaf base, the frequency of cells in the G0/G1 phase of the cell cycle and mitotic activity decreased with leaf age and with the distance from leaf base. No significant differences in 2C nuclear DNA content were found for leaf segments of different age and position. The mean 2C nuclear DNA content was 5.74 pg. The results suggested that the loss of embryogenic competence in mature maize leaf tissues is not related to changes in nuclear DNA content, and that the competence is not simply related to a certain distribution of cells within the cell cycle or to a certain level of mitotic activity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00036227
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  • 6
    Electronic Resource
    Electronic Resource
    Bivariate flow cytometry DNA/BrdUrd analysis of plant cell cycle (1999)
    Springer
    Methods in cell science 21 (1999), S. 155-166 
    Details
    ISSN: 1573-0603
    Keywords: Bromodeoxyuridine ; Immunofluorescence ; Pisum sativum ; Vicia faba ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We describe a protocol for flow cytometry analysis of cell cycle in plants using indirect immunolabelling staining and Vicia faba, Pisum sativum and Zea mays root tip cells as model systems. The protocol is based on simultaneous analysis of two fluorescent signals. The first, obtained after staining with propidium iodide, is used to quantify total nuclear DNA content. The second, obtained after indirect immunofluorescent staining of bromodeoxyuridine (BrdUrd), is used to quantify the amount of BrdUrd incorporated into nuclear DNA. In an attempt to standardize the procedure, the effects of various conditions for partial DNA denaturation using HCl, as well as of BrdUrd concentration and incorporation time on flow cytometry DNA / BrdUrd content analysis have been studied. Maximum BrdUrd-linked fluorescence was observed after a 30 min pulse with 10 μM BrdUrd and after DNA denaturation with 1.5 N HCl (final concentration) for 30 min at 25 °C. Under these conditions, DNA content histograms with relatively small coefficient of variation (〈 4%, full peak) could be obtained. To avoid non-specific staining of cytoplasm and cell walls, the protocol involves the use of nuclei isolated from formaldehyde-fixed tissues. Fixed isolated nuclei are stable and may be stored in hexylene glycol 0.75 M at 4 °C for prolonged periods prior to actual staining and analysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1023/A:1009893008892
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