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  • 1
    ISSN: 1432-2048
    Keywords: Germination (pollen) ; Nicotiana (pollen germination) ; Nucleus (generative, vegetative) ; Pollen(germination, ultrastructure)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The vegetative nucleus (VN) of Nicotiana tabacum L. has been qualitatively and quantitatively studied in fresh, hydrated, and activated pollen. Techniques included the use of optical sectioning by confocal scanning laser microscopy to obtain volume and surface area measurements, and stereoscopic pairs; and freeze-etch electron microscopy to estimate the frequency of nuclear pores per μm2 in the vegetative nucleus. Several morphological changes were observed to occur in pollen grain nuclei during the early processes of tube growth. In freshly dehisced pollen grain, the vegetative and generative nuclei were side by side, but following hydration and activation of the grain, the elongated generative nucleus became partially surrounded by the vegetative nucleus. It was found that during hydration, the surface area of the vegetative nucleus increased and there was a decrease in the frequency of nuclear pores. The calculated total number of pores remained similar. After activation and pollen-tube growth, the vegetative nucleus retained the same surface area as in the hydrated state but the frequency of nuclear pores decreased; therefore, the calculated total number of pores was significantly lowered. When considered alongside complementary biochemical data, these morphological results indicate that RNA production in the vegetative nucleus decreases following germination.
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  • 2
    ISSN: 1432-2048
    Keywords: Arabinogalactan epitope (immunofluores-cence localization) ; Cell wall ; Monoclonal antibody (JIM 8, MAC 207) ; Nicotiana ; Pollen tube
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The monoclonal antibodies MAC 207 and JIM 8, recognizing arabinogalactan epitopes, were used to localize the corresponding antigenic sites in pollen tubes of Nicotiana tabacum L. grown in vitro or semi in vivo. The analysis of the immunofluorescence labelling was performed by means of confocal laser scanning microscopy. Most pollen tubes were labelled along their length, with the exception of the tip region, in a ring-like pattern with remarkable periodicity. The diameter of the rings was approx. 12 μm and the distance between two rings was about 6 μm. No labelling of the cytoplasm, the vegetative nucleus or the generative cell was observed. The presence of labelling in the non-apical tube wall after pectinase and cellulase digestion indicates that the epitopes for MAC 207 and JIM 8 are located in the inner callosic sheath of the pollen-tube wall.
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  • 3
    ISSN: 1432-2048
    Keywords: Cytokinesis ; Generative cell ; Mitosis ; Ornithogalum ; Phragmoplast ; Pollen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The organization of the microtubule (Mt) cytoskeleton during mitosis and cytokinesis of the generative cell (GC) in Ornithogalum virens L. (bicellular pollen type, chromosome number, n = 3) from prophase to telophase/sperm formation was investigated by localization of α-tubulin immunofluorescence using a conventional fluorescence microscope and a confocal laser scanning microscope. Chromosomes were visualized with DNA-binding fluorochrome dyes (ethidium bromide and 4′6-diamino-2-phenyl-indole). The GC of O. virens is characterized by G2/M transition within the pollen grain and not in the pollen tube as occurs in the majority of species with bicellular pollen. It was found that prophase in the GC starts before anthesis and prometaphase takes place after 10 min of pollen germination. The prophase Mts are organized into three prominent bundles, located near the generative nucleus. The number of these Mt bundles is the same as the number of GC chromosomes, a relation which has not previously been considered in other species. The most evident feature in the prophase/ prometaphase transition of O. virens GC is a direct rapid rearrangement of Mt bundles into a network which appears to interact with kinetochores and form a typical prometaphase Mt organization. The metaphase chromosomes are arranged into a conventional equatorial plate, and not in tandem as is thought to be characteristic of GC metaphase. The metaphase spindle consists of kinetochore fibres and a few interzonal fibres which form dispersed poles. Anaphase is characterized by a significant elongation of the mitotic spindle concomitant with the extension of the distance between the opposite poles. At anaphase the diffuse poles converge. Cytokinesis is realized by cell plate formation in the equatorial plane of the GC. The phragmoplast Mts between two future sperm nuclei appear after Mts of the mitotic spindle have disappeared.
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  • 4
    ISSN: 1432-2048
    Keywords: Cell wall (pectin ring) ; Enforced growth fluctuation ; Intracellular free Ca2+ ; Lilium ; Pollentube ; Turgor pressure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Normally growing lily (Lilium longiflorum Thunb.) pollen tubes cultured in standard sucrose medium display a relatively steady tip-growth pattern and a rather even pectin sheath in the cell wall. In an attempt to better understand pulsatory growth, observed in some species, e.g., Petunia, and its possible role in causing the formation of thickened cell wall rings, we have imposed marked fluctuations in the growth-rate of lily pollen tubes. The appropriate growth-perturbing conditions were achieved by modulating the medium osmolarity or by applying caffeine, a non-turgor inhibitor, in a specially designed incubation chamber with a controlled medium flow. The relatively non-esterified pectin deposition in the wall of the growth-interrupted pollen tubes was detected by immunofluorescence microscopy using a monoclonal antibody, JIM 5. The observations show that the periods of slow or inhibited growth correspond to the times when the thickened walls are deposited. Since the growth fluctuations were induced by both turgor- and non-turgor-related means, the proposed endogenous regulatory role of turgor pressure is questioned. Other factors, such as the tip-focused Ca2+ gradient which was demonstrated by ratiometric ion imaging, and the alteration in the extensibility of the cell wall, which correlated with pectin esterification/de-esterification, emerge as candidates for the regulation of growth fluctuations.
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  • 5
    ISSN: 1432-2145
    Keywords: Arabidopsis thaliana ; Meiosis ; Abnormal tetrads ; Male sterility
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A male-sterile mutant, mei-1, of Arabidopsis thaliana is described. In this mutant, instead of a tetrad of four microspores being formed after meiosis, a “tetrad” consisting of from five to eight microspores is formed. The microspores show a wide range of sizes and of DNA contents. The mutant is female-fertile. This mutant was produced by seed transformation with Agrobacterium and appears to be T-DNA tagged.
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  • 6
    ISSN: 1432-2145
    Keywords: Key words Generative cell ; Nicotiana tabacum ; Immunogold ; Myosin ; Pollen tube
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  In this study, polyclonal anti-myosin antibodies were used for immunogold labeling of ultrathin sections of pollen tubes of Nicotiana tabacum L. to unravel the ultrastructural localization of myosin associated with the generative cells. Clusters of immunogold particles were consistently found in association with the area of the outer surface of the vegetative cell plasma membrane present around the generative cell. Compared to the generative cell cytoplasm, the nucleoplasm showed higher numbers of gold particles. This is the first direct evidence demonstrating the presence of myosin in the nuclei of the generative cell of flowering plants. The possible implications of these findings are discussed in relation to movement of the generative cell in the pollen tube cytoplasm.
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 9 (1996), S. 312-317 
    ISSN: 1432-2145
    Keywords: Pollen tube ; Nicotiana tabacum ; Ginkgo biloba ; Dynein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Microtubules in pollen tubes are evident within the vegetative and generative cell cytoplasm. This observation led to the formulation of several hypotheses regarding the role of microtubules in cytoplasmic movement and the migration of the vegetative nucleus/generative cell along the pollen tube. The study of microtubular motor proteins in pollen tubes followed the discovery and characterization of an immunoreactive homolog of mammalian kinesin in tobacco pollen tubes. Recent identification of dynein-related polypeptides in pollen tubes ofNicotiana tabacum and pollen ofGinkgo biloba is a significant step in the definition of the role of microtubule function within pollen and pollen tubes.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 1 (1988), S. 240-247 
    ISSN: 1432-2145
    Keywords: Cell division ; Pollen ; Generative cell ; Sperm cells ; Hordeum vulgare L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Shortly before and during division, the generative cell of barley (Hordeum vulgare L.) is located near the vegetative nucleus, in the peripheral layer of the highly vacuolated vegetative cell at the aperture pole. This position is also characteristic of the two resulting sperm cells. Conventional mitosis of the generative cell is followed by cytokinesis through cell plate formation. Just after division, the two sperm cells are enclosed together within a common “inner” vegetative cell plasma membrane, and they gradually separate from each other only during pollen maturation. The space between the generative or sperm cell plasma membrane and the vegetative cell plasma membrane is very thin and appears to be devoid of a cell wall. Both the generative cell and the young sperm cells contain a normal set of organelles; plastids devoid of starch are only sporadically observed. Our data indicate that in Hordeum vulgare the generative cell divides after migrating inside the pollen grain. This follows the pattern of development well established for several species with tricellular pollen.
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  • 9
    ISSN: 1432-2145
    Keywords: Key words Pollen tube ; Nicotiana tabacum ; Dynein heavy chain related polypeptides ; Immunolocalization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  It is known that pollen tubes contain two high molecular weight polypeptides which share some biochemical and immunological properties with dynein heavy chains. This paper reports data on the subcellular localization of the two dynein heavy chain-related polypeptides during pollen tube growth. Immunofluoresence studies using a purified antibody (Dy-1) raised against a synthetic peptide reproducting the P-loop conserved sequence of dynein heavy chains showed spot-like structures, with a characteristic distribution pattern that depended on the tube length. Biochemical evidence confirmed the presence of dynein heavy chain-related bands in the pollen tube membrane fraction. The association of proteins carrying dynein heavy chain-related polypeptides to cell membranes was affected by detergent (Triton×100), whereas other stripping agents, like NaCl and Na2CO3, did not significantly influence the interaction of dynein heavy chain-related doublet with their cytoplasmic targets. These data suggest that dynein heavy chain-related polypeptides associate with membranous organelles within the vegetative cell of Nicotiana tabacum pollen tubes, implying their involvement in the cytoplasmic distribution of these organelles.
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  • 10
    ISSN: 1432-2145
    Keywords: Key words Freeze-substitution ; Transmission electron microscopy ; Brassica napus ; Tapetum ; Microspore ; Immunogold
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The method of rapid freeze-fixation and substitution was used with Brassica napus floral bud material in order to improve the preservation of microspore and tapetal organelle structure. When observed using transmission electron microscopy, the appearance of the freeze-substituted material differs in a number of ways from the chemically-fixed material previously studied, in particular for the lipid-rich elaioplasts and tapetosomes in the tapetal cells. The tapetosomes have a very electron-dense, opaque appearance when visualized after rapid fixation. In addition, we were able to observe other cytoplasmic details such as pockets in the endoplasmic reticulum and cytoskeletal structures such as microfilaments. Extracellular material was also well-preserved; for example, the fibrous material in the baculae of the developing microspore exine was also visible. Finally, in the freeze-fixed sections specific structures such as elaioplasts could be labelled by antibodies, which indicates that this method preserved protein epitopes that were destroyed by chemical fixation.
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