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  • Pollen  (2)
  • Biochemistry and Biotechnology  (1)
  • Epidermis  (1)
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  • 1
    ISSN: 1432-2048
    Keywords: Brassica ; Embryogenesis ; Heat shock ; Microspore ; Pollen ; Rapeseed
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Elevation of the culture temperature to 32°C for approximately 8 h can irreversibly change the developmental fate of isolated Brassica napus microspores from pollen development to embryogenesis. This stress treatment was accompanied by de-novo synthesis of a number of heat-shock proteins (HSPs) of the 70-kDa class: HSP68 and HSP70. A detailed biochemical and cytological analysis was performed of the HSP68 and HSP70 isoforms. Eight HSP68 isoforms, one of which was induced three fold by the stress treatment, were detected on two-dimensional immunoblots. Immunocytochemistry revealed a co-distribution of HSP68 with DNA-containing organelles, presumably mitochondria. Six HSP70 isoforms were detected, one of which was induced six fold under embryogenic culture conditions. During normal pollen development, HSP70 was localized in the nucleoplasm during the S phase of the cell cycle, and predominantly in the cytoplasm during the remainder. Induction of embryogenic development in late unicellular microspores was accompanied by an intense anti-HSP70 labeling of the nucleoplasm during an elongated S phase. In early bicellular pollen the nucleus of the vegetative cell, which normally does not divide and never expresses HSP70, showed intense labeling of the nucleoplasm with anti-HSP70 after 8 h of culture under embryogenic conditions. These results demonstrate a strong correlation between the phase of the cell cycle, the nuclear localization of HSP70 and the induction of embryogenesis. As temperature stress alone is responsible for the induction of embryogenic development, and causes an altered pattern of cell division, there might be a direct involvement of HSP70 in this process.
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  • 2
    ISSN: 1432-2048
    Keywords: Brassica ; Embryogenesis ; Heat shock ; Microspore ; Pollen ; Rapeseed
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Elevation of the culture temperature to 32°C for approximately 8 h can irreversibly change the developmental fate of isolatedBrassica napus microspores from pollen development to embryogenesis. This stress treatment was accompanied by de-novo synthesis of a number of heat-shock proteins (HSPs) of the 70-kDa class: HSP68 and HSP70. A detailed biochemical and cytological analysis was performed of the HSP68 and HSP70 isoforms. Eight HSP68 isoforms, one of which was induced three fold by the stress treatment, were detected on two-dimensional immunoblots. Immunocytochemistry revealed a co-distribution of HSP68 with DNA-containing organelles, presumably mitochondria. Six HSP70 isoforms were detected, one of which was induced six fold under embryogenic culture conditions. During normal pollen development, HSP70 was localized in the nucleoplasm during the S phase of the cell cycle, and predominantly in the cytoplasm during the remainder. Induction of embryogenic development in late unicellular microspores was accompanied by an intense anti-HSP70 labeling of the nucleoplasm during an elongated S phase. In early bicellular pollen the nucleus of the vegetative cell, which normally does not divide and never expresses HSP70, showed intense labeling of the nucleoplasm with anti-HSP70 after 8 h of culture under embryogenic conditions. These results demonstrate a strong correlation between the phase of the cell cycle, the nuclear localization of HSP70 and the induction of embryogenesis. As temperature stress alone is responsible for the induction of embryogenic development, and causes an altered pattern of cell division, there might be a direct involvement of HSP70 in this process.
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  • 3
    ISSN: 1615-6102
    Keywords: Helianthus annuus ; Embryo ; Epidermis ; Protoderm ; Phenylalkylamine ; Ion channel
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A fluorescently labeled phenylalkylamine, DM-Bodipy PAA, was used as a probe for the in vivo detection of ion channels in embryonic and nonembryonic tissues of sunflower. Zygotic embryos, somatic embryos, callus, leaves, roots, and shoots were analysed. Fluorescence intensity in the tissues was determined with cytofluorometry and confocal microscopy. DM-Bodipy PAA intensively labeled the protoderm and epidermis cells in both zygotic and somatic embryos. Callus cultures exhibited labeling on sites where somatic embryos developed. Labeling was, however, very weak in leaves, shoots, and roots, except in the root cap and in the epidermis of the root. Considering that the location of phenylalkylamine binding sites is related to the distribution of ion channels in both animal and plant cells, the high intensity of labeling observed in the protoderm and epidermis of zygotic and somatic embryos as well as in protoderm, epidermis, and caps of root tips, is consistent with the role these tissues may play in ion exchange with the environment.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 55 (1997), S. 630-641 
    ISSN: 0006-3592
    Keywords: nitrification ; immobilized cells ; Nitrosomonas europaea ; substrate limitation ; biomass death ; staining techniques ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: The dynamics of growth and death of immobilized Nitrosomonas europaea were studied. For this, the death rate of suspended cells was determined in the absence of ammonium or oxygen by following the loss of respiration activity and by fluorescein-diacetate (FDA)/lissamine-green staining techniques. The death rates obtained (1.06 × 10-6 s-1 or 4.97 × 10-6 s-1 in the absence of oxygen or ammonium, respectively) were incorporated in a dynamic growth model and the effects on the performance of the immobilized-cell process illustrated by model simulations.These model simulations and experimental validation show that if decay of biomass occurs the biomass concentration in the center of the bead decreases. As a result, the systems react slower to changes in substrate concentrations than if all cells remain viable.To show that cells in the center of the bead died, the FDA and lissamine-green staining techniques were adapted for immobilized cells. It was shown that biomass decay occurred, especially in the center of the bead; the amount of cells decreased there, and the remaining cells were all stained with lissamine green indicating cell death. After the substrate availability was decreased, also cells near the surface of the bead lost their viability. The number of viable cells increased again after increasing the substrate concentration as the result of cell multiplication. At low substrate concentrations and low hydraulic retention times, as for example in the treatment of domestic wastewater, the death rate of cells is thus an important parameter for the performance of the immobilized-cell system. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 630-641, 1997.
    Additional Material: 6 Ill.
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