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  • 1995-1999  (2)
  • Freeze substitution  (1)
  • Nicotiana tabacum
  • 1
    ISSN: 1615-6102
    Keywords: Caffeine ; Freeze substitution ; Lilium ; Pollen tubes ; Rapid freeze fixation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In an attempt to correlate structural effects with the known dissipation of the tip-focused Ca2+ gradient caused by caffeine, we have examined the ultrastructure of caffeine-treated lily pollen tubes prepared by rapid freeze fixation and freeze substitution. We show that treatment with caffeine results in a rapid rearrangement of secretory vesicles at the pollen tube tip; the normal cone-shaped array of vesicles is rapidly dispersed. In addition, microfilament bundles appear in the tip region, where they had previously been excluded. Delocalized vesicle fusion continues in the presence of caffeine but tube extension ceases. Removal of caffeine from the growth medium initially causes tip swelling, delocalized vesicle fusion and presence of microfilaments well into the tip before normal structure and growth resume, concurrent with the previously reported return to a normal Ca2+ gradient.
    Type of Medium: Electronic Resource
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  • 2
    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
    Library Location Call Number Volume/Issue/Year Availability
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