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  • 1
    Electronic Resource
    Electronic Resource
    Distribution of microtubules during the growth of tobacco pollen tubes
    Amsterdam : Elsevier
    Biology of the Cell 79 (1993), S. 125-132 
    Details
    ISSN: 0248-4900
    Keywords: Nicotiana tabacum ; acetylated α-tubulin ; confocal laser scanning microscope ; microtubules ; pollen tube growth ; tyrosinated α-tubulin
    Source: Elsevier Journal Backfiles on ScienceDirect 1907 - 2002
    Topics: Biology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/0248-4900(93)90248-D
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  • 2
    Electronic Resource
    Electronic Resource
    In vitro double fertilization in Nicotiana tabacum (L.): polygamy compared with selected single pair somatic protoplast and chloroplast fusions (2000)
    Springer
    Sexual plant reproduction 13 (2000), S. 113-117 
    Details
    ISSN: 1432-2145
    Keywords: Key words Cell fusion ; Gamete interaction ; In vitro polygamy ; Nicotiana tabacum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  In vitro polygamy was studied mainly by using isolated sperm and central cells of tobacco in order to elucidate the mechanism that might be involved in preventing in vivo polygamy. In 17.5% 4000 M.W. polyethylene glycol, only when two sperm cells were made close enough to each other and adhered to a female cell simultaneously was polygamy possible. If one sperm cell fused with the egg or central cell, within 30 min another sperm cell could not fuse with the same egg or central cell. Similar phenomena were found in selected single somatic cell fusion. When more than two protoplasts adhered to each other simultaneously, fusion was always successful; after two protoplasts fused, within 30 min the fusion products could not fuse with another protoplast under the same conditions. This comparative study revealed this characteristic to be shared by both sexual and somatic cell fusion. However, after cytoplasm reorganization was complete in the fusion product, it was possible for the fusion product to fuse with the third protoplast. This indicates that the obstruction to additional fusion was present only during a certain period after the preceding fusion under certain condition. The possible reason for the effect is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004970000044
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  • 3
    Electronic Resource
    Electronic Resource
    Responses of tobacco pollen to high humidity and heat stress: viability and germinability in vitro and in vivo (1991)
    Springer
    Sexual plant reproduction 4 (1991), S. 104-109 
    Details
    ISSN: 1432-2145
    Keywords: High humidity and temperature stress ; Nicotiana tabacum ; Tobacco ; Pollen viability ; Vigour ; Semi-vivo technique
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Responses of pollen grains of Nicotiana tabacum to high humidity (95% RH, 4 h) and temperature (38°/45° C, 4 h) stresses were investigated. Pollen grains were subjected to only RH or only temperature, or to both of these stresses. Their viability was assessed on the basis of the fluorochromatic reaction (FCR) test, and vigour was assessed on the basis of the time taken for in vitro germination as well as on the emergence of pollen tubes through the cut end of semi-vivo implanted styles. None of the stress conditions affected pollen viability and high RH or high temperature stress did not individually affect pollen vigour. However, pollen vigour was markedly affected when both the stresses were given together. Pollen grains subjected to high RH at 38° C took a longer time to germinate in vitro and the pollen tubes emerged later from the cut end of the semi-vivo styles; division of the generative cell was also delayed. Pollen grains subjected to high RH at 45° C failed to germinate in vitro, but did germinate on the stigma. Many pollen tubes subjected to this treatment showed abnormalities, and the growth of pollen tubes in the pistil was much slower than that observed in other treatments. Pollen samples subjected to all of the stress conditions were able to induce fruit and seed set. The implications of these results on the relationship between the FCR test and viability, and between viability and vigour, especially in stressed pollen, are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00196495
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  • 4
    Electronic Resource
    Electronic Resource
    The microtubule cytoskeleton and the rounding of isolated generative cells ofNicotiana tabacum (1992)
    Springer
    Sexual plant reproduction 5 (1992), S. 64-71 
    Details
    ISSN: 1432-2145
    Keywords: Generative cell ; Isolation ; Microtubules ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Upon squashing of the pollen grain, the isolated generative cell ofNicotiana tabacum looses its spindle shape to become spherical; this phenomenon is independent of the sucrose concentration used. The time necessary for this change can vary from 1 min (0% sucrose) to 20 min (30% sucrose). The microtubular cytoskeleton was studied by means of immunofluorescence and electron microscopy. Just after isolation, 5 to 15 clearly visible bundles in microtubules organized in a basket-like structure are present. After 15 min in medium with 15% sucrose, the microtubular cytoskeleton disappears, and a diffusely spread tubulin can be observed. Neither the addition of 10–20 μM taxol to the medium, nor the omission of Ca2+ to the medium has any effect on the changes in cell shape and loss of microtubular bundles after isolation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00714559
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  • 5
    Electronic Resource
    Electronic Resource
    Effects of high humidity and temperature stress on pollen membrane integrity and pollen vigour in Nicotiana tabacum (1989)
    Springer
    Sexual plant reproduction 2 (1989), S. 137-141 
    Details
    ISSN: 1432-2145
    Keywords: Humidity ; Temperature stress ; Nicotiana tabacum ; Pollen germination ; Pollen membrane
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The prolonged exposure of pollen Nicotiana tabacum to high humidity at both room temperature and 38° C did not affect membrane integrity as revealed by the fluorochromatic reaction (FCR) test, but did affect pollen vigour. At room temperature germination was not affected, although tube growth was reduced; at 38° C, there was both a reduction in tube growth and delayed germination. When the pollen was subjected to 1 h hydration followed by 1 h desiccation (up to a maximum of four cycles) at room temperature, a reduction in the FCR, germination and tube length after each desiccation treatment was observed. Subsequent hydration fully restored the FCR, but only partially restored germination and tube growth. At 38° C, however, FCR, germination, and tube growth were drastically reduced. The implications of these results on the relationship between FCR and germinability, the responses of pollen exposed to humidity and temperature stress in the field, and on pollen storage are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00192759
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  • 6
    Electronic Resource
    Electronic Resource
    Cytoskeletal changes during generative cell division and sperm formation inTradescantia virginiana (1989)
    Springer
    Protoplasma 150 (1989), S. 54-71 
    Details
    ISSN: 1615-6102
    Keywords: Generative cell ; Microtubules ; Mitosis ; Cytokinesis ; Pollen ; Sperm ; Tradescantia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cytoskeletal organization and chromosome behavior were studied inTradescantia generative cells prior to and during sperm formation using in vitro grown pollen tubes and fluorescence staining methods. Before pollen germination, the crescent-shaped generative cell contains a reticulate microtubule (Mt) system. The cell elongates dramatically after germination, and its Mts assume a helical to longitudinal arrangement. Chromosome condensation is evident approximately 3hr after germination. Kinetochores appear as dark interruptions in the Mt array, and thus seem to attach directly to interphase fibers. No metaphase plate typical of other cells is observed with either DAPI or anti-tubulin staining. Instead, the chromosomes adopt a twisted or braided arrangement, with kinetochores distributed along the length of the cell and kinetochore fibers linked to each other and to surrounding fibers. Anaphase is characterized by a staggered, overlapping separation of chromosomes and by elongation of Mt branches connecting opposing kinetochore fibers. Cytokinesis appears to utilize a furrowing process; a phragmoplast or cell plate was never seen. As a result of these events, the sperm directly inherit their cytoskeleton from generative cell Mts involved in division. No actin fibers are observed at any stage using rhodamine-phalloidin staining. The results are discussed in terms of other reports on sperm formation, possible mitotic and cytokinetic mechanisms, and past distinctions between Mt arrays in higher plant somatic cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01352921
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  • 7
    Electronic Resource
    Electronic Resource
    The organization of the cytoskeleton in the generative cell and sperms ofHyacinthus orientalis (1992)
    Springer
    Protoplasma 168 (1992), S. 41-50 
    Details
    ISSN: 1615-6102
    Keywords: Pollen tube ; Microtubules ; Cellular division ; Generative cell ; Sperm cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The microtubular cytoskeleton of the generative cell (GC) ofHyacinthus orientalis has been studied until the formation of the sperm cells (SCs). Immunofluorescence procedures in combination with confocal laser scanning microscopy (CLSM) has enabled the visualization of the organization of the microtubular cytoskeleton. Chemical fixation and freeze-fixation electron microscopy have been used to investigate the cytoskeleton and the ultrastructural organization of the GC and SCs. During pollen activation the GC is spindle-shaped. Microtubules (MTs) are organized as bundles and distributed in proximity of the GC plasmamembrane, forming a basket-like structure. Following migration through the pollen tube, the basket-like structure becomes more intertwined. During the nuclear division the MTs are involved in the segregation of the chromosomes and kinetochores are clearly discernible. Association with organelles is also observed. The chromosomes of the GC remain condensed until they separate in two sperm nuclei. The pre-prophase band was never observed. At the end of the GC division the microtubular network reorganizes in the two SCs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01332649
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  • 8
    Electronic Resource
    Electronic Resource
    The organization of microtubules during generative-cell division inConvallaria majalis (1999)
    Springer
    Protoplasma 207 (1999), S. 147-153 
    Details
    ISSN: 1615-6102
    Keywords: Cell division ; Confocal microscopy ; Convallaria majalis ; Generative cell ; Liliaceae ; Microtubules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The organization of the microtubule cytoskeleton in the generative cell ofConvallaria majalis has been studied during migration of the cell through the pollen tube and its division into the two sperm cells. Analysis by conventional or confocal laser scanning microscopy after tubulin staining was used to investigate changes of the microtubule cytoskeleton during generative-cell migration and division in the pollen tube. Staining of DNA with 4′,6-diamidino-2-phenylindole was used to correlate the rearrangement of microtubules with nuclear division during sperm cell formation. Before pollen germination the generative cell is spindle-shaped, with microtubules organized in bundles and distributed in the cell cortex to form a basketlike structure beneath the generative-cell plasma membrane. During generative-cell migration through the pollen tube, the organization of the microtubule bundles changes following nuclear division. A typical metaphase plate is not usually formed. The generative-cell division is characterized by the extension of microtubules concomitant with a significant cell elongation. After karyokinesis, microtubule bundles reorganize to form a phragmoplast between the two sperm nuclei. The microtubule organization during generative-cell division inConvallaria majalis shows some similarities but also differences to that in other members of the Liliaceae.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01282994
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  • 9
    Electronic Resource
    Electronic Resource
    The anti-centrosome monoclonal antibody 6C6 reacts with a plasma membrane-associated polypeptide of 77 kDa fromNicotiana tabacum pollen tubes (1996)
    Springer
    Protoplasma 190 (1996), S. 68-78 
    Details
    ISSN: 1615-6102
    Keywords: Microtubule ; Microtubule organizing centers ; Nicotiana tabacum ; Pericentriolar antigens ; Plasma membrane ; Pollen tube
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In the pollen and pollen tube of higher plants, the distribution of the microtubular cytoskeleton has been extensively studied. Even though the pattern of microtubules is known, one of the most remarkable deficiencies is the absence of data on the localization of microtubule-nucleation sites in the pollen tubes. In order to get insights about the localization of centrosome-like structures in the pollen tube ofNicotiana tabacum L., we have used the monoclonal antibody 6C6 to search for pericentriolar antigen(s). The antibody was initially raised against a component of animal centrosomes and has been already employed to locate centrosomal structures in other plant cell types. By immunoblotting analysis, a polypeptide of Mr 77,000 was identified specifically in the membrane-associated protein fraction of the pollen tube, and is absent from the soluble protein pool. Immunofluorescence observations have shown the polypeptide to be located in the apical part of the pollen tube (about 40–50 μm from the tip) in association with the cortical area. A purified plasma membrane fraction from the growing pollen tubes has been obtained, using H+-ATPase activity as an organelle marker. The plasma membrane fraction was shown to be enriched in the Mr 77,000 polypeptide, which can be extracted from membranes by treatment with the detergent CHAPS at a concentration of 0.5%. These data open new research perspectives on the localization and analysis of putative cortical microtubule nucleation sites in the pollen tube.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01281195
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