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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 210 (2000), S. 188-201 
    ISSN: 1615-6102
    Keywords: Microtubules ; Oryza sativa ; Rice ; Microspore formation ; Pollen development ; Asymmetrical cell division
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Anthers of rice (Oryza sativa L.) at different stages of development were cryofixed, freeze-substituted, and embedded in methacrylate. Sections were then cut and immuno-labeled with anti-tubulin to localize microspore microtubules. Changes in microtubule distribution pattern were followed by confocal fluorescence microscopy. To facilitate description, pollen development has been divided into four developmental stages (twenty-four phases). (i) The young-microspore stage (phases 1–5) is characterized by the formation of a vacuole. When the vacuole enlarges, the nucleus moves to the periphery of the cell. Afterwards the nucleus migrates to a site opposite the germ pore. Between the germ pore and the nucleus there is a strand of cytoplasm which contains some microtubules that run in parallel to the pore-nucleus axis. (ii) At the first-mitosis stage (phases 6–12) a perinuclear band of tubules appears which eventually girdles the nucleus. (iii) At the generative-cell development stage (phases 13–20), after the first mitosis a generative cell forms at a site opposite the germ pore. It is initially lens-shaped and part of its wall is appressed closely to the plasma membrane of the microspore. The wall of the generative cell contains cellulose and callose. Later the generative cell detaches from the microspore wall and migrates into the cytoplasm of the vegetative cell. In the vegetative cytoplasm the generative cell becomes spherical. (iv) At the second-mitosis and sperm formation stage (phases 21–24) the mitotic division is symmetrical. Before division, the shape of the generative cell changes from spherical to spindle-shaped. After cell division two sperm cells form, although they do not detach from each other. Later, cytoplasmic processes containing microtubules develop at the two ends of the sperm cells. These long processes remain linked to each other to form a paired unit.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 15 (1996), S. 396-400 
    ISSN: 1432-203X
    Keywords: Brassica campestris ssp. parachinensis ; Microspore ; Embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Microspores of several genotypes of Brassica campestris ssp. parachinensis have been cultured in vitro and induced to undergo embryogenesis and plant formation. Conditions favourable for embryogenesis in this species include a bud size of 2–2.9 mm, NLN-13 culture medium (Nitsch and Nitsch 1967; Lichter 1981, 1982; Swanson 1990), and an induction through exposure to 32°C for a period of 48 h. Longer periods of an elevated temperature for induction of embryogenesis resulted in embryo abortion at early developmental stages. With the protocol developed here, microspores of 60–80% of donor plants could be induced to produce embryos, although embryo yields were low, i.e. 2–5 embryos per 10 buds. Some genotypes responded to culture conditions with high numbers of embryo formation (100–150 embryos per 10 buds) but most of these subsequently failed to mature. The pattern of cell division and morphological changes of the microspores in culture were studied using various microscopic techniques.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 5 (1992), S. 182-188 
    ISSN: 1432-2145
    Keywords: Isolated generative cells ; Microtubules ; Mitosis ; Immunofluorescence ; Confocal microscope ; Allemanda neriifolia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Organizational changes in the microtubules of isolated generative cells of Allemanda neriifolia during mitosis were examined using anti-α-tubulin and confocal laser scanning microscopy. Due to an improved resolution and a lack of out-of-focus interference, the images of the mitotic cytoskeleton obtained using the confocal microscope are much clearer than those obtained using the non-confocal fluorescence systems. In the confocal microscope one can see clearly that the spindle-shaped interphase cells contain a cage-like cytoskeleton consisting of numerous longitudinally oriented microtubule bundles and some associated smaller bundles. At prophase, the shape of the cells invariably becomes spherical. The microtubule cytoskeleton inside the cells concomitantly changes into a less organized form — consisting of thick bundles, patches, and dots. This structural form is not very stable, and soon afterwards the cytoskeleton changes into a reticulate network. Then the nuclear envelope breaks down, and the microtubules become randomly dispersed throughout the cell. Afterwards, the microtubules reorganize themselves into a number of half-spindle-like structures, each possessing a microtubule-nucleating center. The locations of these centres mark out the positions of the presumptive spindle poles. Numerous microtubules radiate from these centres toward the opposite pole. At metaphase, the microtubules form a number of bipolar spindles. Each spindle has two half-spindles, and each half-spindle has a sharply focused microtubule centre at the pole region. From the centres, kinetochore and non-kinetochore microtubules radiate toward the opposite half-spindle. At anaphase A, sister chromatids separate, the cells elongate, and the kinetochore microtubules disappear; the non-kinetochore microtubules, however, remain, and a new array of microtubules, in the form of a cage, appears. The peripheral cage bundles and the non-kinetochore bundles coverge into a sharp point at the pole region. Later, at anaphase B the microtubule cytoskeleton undergoes reorganization giving rise to a new array of longitudinally oriented microtubule bundles in the cell centre and a cage-like cytoskeleton in the periphery. At telophase, some of the cells elongate further, but some become spherical. The microtubules in the central region of the elongated cell become partially disrupted due to the formation of a phragmoplast-junction-like structure in the mid-interzone region. The microtubule bundles at the periphery are spirally organized, and they appear not to be disrupted by the phragmoplast-like junction. The microtubules in the spherical telophase cells (unlike those seen in the elongated telophase cells) are arranged differently, and no phragmoplast-junction-like structure forms in the spherical cells. The structural and functional significances of some of these new features of the organization of the microtubule cytoskeleton as revealed by the confocal microscope are discussed.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 5 (1992), S. 27-33 
    ISSN: 1432-2145
    Keywords: Isolated generative cells ; Ultrastructure ; Microtubule ; Immunofluorescence microscopy ; Allemanda neriifolia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ultrastructure of isolated generative cells ofAllemanda neriifolia at interphase and prophase was studied. The microtubule organization of the isolated cells was also investigated by immunofluorescence microscopy with a monoclonal anti-α-tubulin. After the generative cells had been isolated from the growing pollen tubes by osmotic shock, most of the cells were at prophase and only a few were at interphase. The interphase cell is spindle shaped and contains an ellipsoidal nucleus. In addition to the usual organelles, the cytoplasm of the interphase cell contains numerous vesicles (each measuring 40–50 nm in diameter) and two sets of longitudinally oriented microtubule bundles — one in the cortical region and the other near the nucleus. Most of the prophase cells are spherical in shape. Based on the ultrastructure and the pattern of microtubule cytoskeleton organization three types of prophase cells can be recognized. (1) Early prophase cell, which contains the usual organelles, numerous vesicles, and a spherical nucleus with condensed chromosomes. Longitudinally oriented microtubule bundles can no longer be seen present in the early prophase cell. A new type of structure resembling a microtubule aggregate appears in the cytoplasm. (2) Mid prophase cell, which has a spherical nucleus containing chromosomes that appear more condensed than those seen in the early prophase cell. In addition to containing the usual organelles, the cytoplasm of this cell contains numerous apparently randomly oriented microtubules. Few vesicles are seen and microtubule aggregates are no longer present. (3) Late prophase cell, typified by the lack of a nuclear envelope. Consequently, the chromosomes become randomly scattered in the cytoplasm. Microtubules are still present and some become closely associated with the chromosomes. The changes in the ultrastructure and in the pattern of microtubule organization in the interphase and prophase cells are discussed in relation to the method of isolation of the generative cells.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 4 (1991), S. 132-137 
    ISSN: 1432-2145
    Keywords: Isolated generative cells ; Microtubules ; Mitosis ; Immunofluorescence ; Allemanda neriifolia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The organizational changes of the microtubules of isolated generative cells of Allemanda neriifolia during division were followed using anti-α-tubulin and immunofluorescence microscopy. Generative cells were isolated from the pollen tubes after osmotic shock treatment. Immediately after isolation most of the cells remain either in early or late prophase. The shape of the cell changes from spindle to spheroidal. In early prophase the nuclear membrane of the cell appears intact and the cytoplasm full of reticulate microtubules of different shapes and thicknesses. Later, the nuclear membrane breaks up. After the nuclear membrane has broken up, the chromosomes scatter into the cytoplasm and mix with the microtubules. When cells enter metaphase, spindle microtubules form. Afterwards, in anaphase, sister chromatids separate and the spindle disappears. A new array of longitudinally oriented cage microtubules appears. As the cells enter early telophase, the cage microtubules disappear and an array of interpolar microtubules begins to form. Later, in some telophase cells the interpolar microtubules become highly elongated, but in others they soon disappear and become replaced by a thick band(s) (or sheet(s)) of microtubules in the midplane between the two clusters of chromosomes and the cell shape reverts back to spheroidal. In culture no phragmoplast junctions appear in any of the late telophase cells although they are present under the in situ condition (i.e. in pollen tubes).
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 3 (1990), S. 213-218 
    ISSN: 1432-2145
    Keywords: Generative cell ; Isolation ; Microtubule ; Immunofluorescence microscopy ; Zephyranthes grandiflora
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Microtubule organization in the generative cells of Zephyranthes grandiflora was investigated by immunofluorescence microscopy with a monoclonal anti-α-tubulin. The experimental materials used were generative cells located within pollen grains and tubes (i.e., in situ) as well as those artificially isolated after osmotic shock or grinding treatments of the pollen grains. Diverse microtubule organization patterns were revealed. In situ, the generative cells appeared spindle-shaped and contained mainly longitudinally oriented microtubule bundles, although other types were found as well. After isolation, as the alteration in microtubule patterns took place, the spindle-shaped generative cells became ellipsoidal and then spherical. In the ellipsoidal cells a transitional form consisting of a mixture of microtubule bundles and meshes could be found. In spherical cells the mesh structure appeared to be the predominant pattern. These results indicate that the microtubule cytoskeleton of the generative cells can change easily from one structural form to another in accordance with environmental conditions and may play an important role in determining the cell shape.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 185 (1995), S. 170-177 
    ISSN: 1615-6102
    Keywords: Bisporic embryo sac ; Cymbidium sinense ; Megasporogenesis ; Meiosis ; Microtubules
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary InCymbidium sinense embryo sac development follows the bisporic pattern. This pattern of development is rare in orchids. Changes in the structure and organization of the microtubular cytoskeleton during megasporogenesis in this orchid were followed using an immunofluorescence technique and confocal microscopy. At the initial stage of development the microtubules in the archespore are randomly oriented throughout the cortex and cytoplasm. Later, microtubules undergo reorganization with more microtubules distributed in the cortex and around the nucleus. Megasporocyte formation is marked by the elongation of the archesporial cell. The cytoskeleton in the elongated megasporocyte is radially organized with microtubules extending from the nucleus to the peripheral region of the cell. In the megasporocyte some of the microtubules also show a polarized pattern of distribution with more microtubules at the chalazal end than the micropylar end. After meiosis I, a dyad is formed. Each dyad member possesses a complement of randomly oriented microtubules. But some microtubules in the dyad also show radial orientation. Soon afterwards microtubules in the degenerating micropylar dyad cell disappear. The functional (chalazal) dyad cell proceeds to meiosis II giving rise to a two-nucleate embryo sac. The pattern of microtubule organization in the two-nucleate embryo sac is quite complex. Each nucleus is surrounded by a set of randomly distributed microtubules. But the vacuolar region between the two nuclei is surrounded by a parallel array of cytoplasmic microtubules. No preprophase bands were observed during the development of the megasporocyte. This is the first description of microtubules in bisporic embryo sac development.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 107 (1981), S. 21-26 
    ISSN: 1615-6102
    Keywords: Adventive embryos ; Coriander ; Expiants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The petiole expiants of coriander when cultured on a solid or liquid Murashige and Skoog's medium supplemented with 0.5 mg/l 2,4-D developed a large number of meristemoids. These meristemoids could then be induced to form adventive embryos when transferred into a liquid MS-medium without 2,4-D. The development of the expiants leading to the formation of meristemoids was followed anatomically and by using SDS-polyacrylamide gel electrophoresis of soluble proteins at various days of culture.
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 170 (1992), S. 86-89 
    ISSN: 1615-6102
    Keywords: Pea ; Allamanda schottii ; Chromosome ; Cytoskeleton
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cytoskeletons surrounding the chromosomes of the root tip cells ofPisum sativum and the generative cells ofAllamanda schottii were visualized using Triton X-100 extraction and scanning electron microscopy. The cytoskeleton surrounding the chromosome consisted of a reticulate network of fibres. This is the first report showing the existence of a “chromosome exo-skeleton” in plant cells.
    Type of Medium: Electronic Resource
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