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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 80 (1990), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: The organization, isolation and physiology of the angiosperm male gamete has recently emerged as an area of special interest. Detailed in vivo studies have revealed: (1) that the functional unit of male reproduction is a‘male germ unit,’composed of two sperm cells and a physically associated vegetative nucleus; (2) that the two sperm cells are often cytoplasmically dimorphic, and (3) that some sperm cells appear to undergo preferential fertilization, preferentially fusing with either the egg or the proendospermaric central cell. Male gamete isolation has provided enrichments of up to 107 cells ml–1, with sperm integrity and viability demonstrated by the flunrochromatic reaction and ATP levels. Although results from biochemical and hybridoma antibody characterization are still preliminary, they indicate that sperm cells possess polypeptide expression patterns that are different from those of surrounding cells. These findings suggest the existence of an independent developmental program in the angiosperm male gamete.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 4 (1991), S. 28-35 
    ISSN: 1432-2145
    Keywords: Tapetal cells ; Brassica oleracea L ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The ultrastructure of the secretory, binucleate tapetum of Brassica oleracea in the micro spore mother cell (MMC) stage through to the mature pollen stage is reported. The tapetal cells differentiate as highly specialized cells whose development is involved in lipid accumulation in their final stage. They start breaking down just before anther dehiscence. Nuclei with dispersed chromatin, large nucleoli and many ribosomes in the cytoplasm characterize the tapetal cells. The wall-bearing tapetum phase ends at the tetrade stage. The dissolution of tapetal walls begins from the inner tangential wall oriented towards the loculus and proceeds gradually along the radial walls to the outer tangential one. The plasmodesmata transversing the radial walls between tapetal cells persist until the mature microspore, long after loss of the inner tangential wall. After wall dissolution, the tapetal protoplasts retain their integrity and position within the anther locule. The tapetal cell membrane is in direct contact with the exine of the microspores/pollen grains and forms tubular evaginations that increase its surface area and appear to be involved in the translocation of solutes from the tapetal cells to the microspores/ pollen grains. The tapetal cells exhibit a polarity expressed by spatial differentiation in the radial direction.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 4 (1991), S. 104-109 
    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
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 4 (1991), S. 145-154 
    ISSN: 1432-2145
    Keywords: Angiosperm gametes ; Egg cell ; Embryosac ; Gamete isolation ; Gametophytic cells ; Sperm cells ; Generative cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The availability of generative cells, sperm cells, embryo sacs and egg cells from angiosperm plants in isolated conditions has opened up many prospects: study of the mechanism of recognition and fusion between gametes of opposite sex and detailed observation of the process of fertilization, biochemical and genetic analysis of gamete-specific components and genetic engineering in combination with in vitro fertilization. This review provides a list of about ninety publications, in which the isolation of male or female angiosperm gametes and the closely related generative cells and embryo sacs is reported. The species used are summarized in two tables. A description is given of the diverse isolation techniques, which consist of enzymatic digestion, bursting of pollen by osmotic shock, squashing, grinding and micro-dissection. Viability of isolated cells and yield, two important aspects of biotechnological manipulation, are emphasized. A critical evaluation of the most significant results obtained so far with isolated material is presented together with notes on prospects for the future.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Sexual plant reproduction 4 (1991), S. 176-181 
    ISSN: 1432-2145
    Keywords: Pollen ; Brassica napus ; Mitoses ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Brassica napus pollen development during the formation of the generative cell and sperm cells is analysed with light and electron microscopy. The generative cell is formed as a small lenticular cell attached to the intine, as a result of the unequal first mitosis. After detaching itself from the intine, the generative cell becomes spherical, and its wall morphology changes. Simultaneously, the vegetative nucleus enlarges, becomes euchromatic and forms a large nucleolus. In addition, the cytoplasm of the vegetative cell develops a complex ultrastructure that is characterized by an extensive RER organized in stacks, numerous dictyosomes and Golgi vesicles and a large quantity of lipid bodies. Microbodies, which are present at the mature stage, are not yet formed. The generative cell undergoes an equal division which results in two spindle-shaped sperm cells. This cell division occurs through the concerted action of cell constriction and cell plate formation. The two sperm cells remain enveloped within one continuous vegetative plasma membrane. One sperm cell becomes anchored onto the vegetative nucleus by a long extension enclosed within a deep invagination of the vegetative nucleus. Plastid inheritance appears to be strictly maternal since the sperm cells do not contain plastids; plastids are excluded from the generative cell even in the first mitosis.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-2145
    Keywords: Female germ unit ; Nicotiana ; Megagametophyte ; Organelle movement ; Video microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Living embryo sacs and megagametophytic cells of Nicotiana alata and Nicotiana tabacum were obtained using enzymatic maceration and microdissection. The yields of isolated embryo sacs, egg apparatus and central cells were up to 35%, 40% and 35%, respectively. Vectorial movement of organelles and undulations of tubular structures, presumably endoplasmic reticulum, were observed in eggs, synergids and central cells using video-enhanced microscopy. Despite evident viability using the fluorochromatic reaction, the egg displays much less organelle movement and therefore appears to be quiescent. The large vacuole of the central cell is traversed by mobile strands of cytoplasm through which organelles migrate. A polygonal network is located at the periphery of the central cell, which may contribute to anchorage of the cell with the embryo-sac wall. The observation of organelle movement provides direct evidence of the condition of the cell and may be a useful approach for assessing cell vigor.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1432-2145
    Keywords: Immunolocalization ; Pistil protein (Sk2) ; Solanum tuberosum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sk2 protein is the most abundant member of the pistil-specific proteins of Solanum tuberosum. Sk2 protein has been localized by use of a polyclonal antibody (anti-Sk2) in the pistils of four clones of Solanum tuberosum. In the stigmas Sk2 protein accumulates to a high level in the cytoplasm of the internal secretory cells underlying the papillae one day prior to anthesis. In styles, the intercellular matrix of the transmitting tissue cells is intensely labelled by anti-Sk2. Sk2 protein is present in all four clones and shows the same labelling pattern. The possible role of the Sk2 protein in pollen tube growth is discussed.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1432-2048
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The development of characteristic cytoplasmic inclusions from endoplasmic-reticulum (ER) cisternae in nucellar cells of some species of Euphorbia has been studied by electron microscopy. The formation of these organelles is preceded by the appearance of rough ER cisternae filled with an electron-dense material and forming complicated networks. Vesicular structures are formed which grow rapidly to give electron-dense, spherical dilations. On the outer surface of their limiting membrane numerous ribosomes and often polysomes are present. This membrane can be seen to remain continuous with the membranes of one or more cisternae of the rough ER up to when the dilations have a maximum diameter of 2.5–3 μ. At this time, continuity between the ER cisternae and the spherical dilations ceases. After this the new cell organelles remain unchanged in size, shape, and electron-density until the cell is disintegrated by the growing embryo-sac. The fate of the contents of these organelles is discussed.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1432-2048
    Keywords: Kinesin ; Nicotiana ; Organelle movement ; Pollen tube
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In plant cells, microtubule-based motor proteins have not been characterized to the same degree as in animal cells; therefore, it is not yet clear whether the movement of organelles and vesicles is also dependent on the microtubular cytoskeleton. In this work the kinesinimmunoreactive homologue from pollen tubes of Nicotiana tabacum L. has been purified and biochemically characterized. The protein preparation mainly contained a polypeptide with a relative molecular weight of approx. 100 kDa. This polypeptide bound to animal microtubules in an ATP-dependent manner and it further copurified with an ATPase activity fourfold-stimulated by the presence of microtubules. In addition, the sedimentation coefficient (approx. 9S) was similar to those previously shown for other kinesins. Immunofluorescence analyses revealed a partial co-distribution of the protein with microtubules in the pollen tube. These data clearly indicate that several properties of the kinesin-immunoreactive homologue are similar to those of kinesin proteins, and suggest that molecular mechanisms analogous to those of animal cells may drive the microtubule-based motility of organelles and vesicles in plants.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1432-2242
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cytological and genetical analyses were made of the breeding system of embryo-cultured interspecific tomato hybrids between L. esculentum and L. peruvianum. It was found that fluorescence techniques and electron microscopy allowed a distinction to be made between pollen tubes inhibited by a unilateral incompatibility reaction and pollen tubes inhibited by a self-incompatibility reaction, after self-pollination of the hybrids or after reciprocal crossing between the hybrid and the parental species. The observed differences, if real and reliable, demonstrate that unilateral incompatibility in esculentum pollen tubes is governed by a single gametophytic factor which is either linked or allelic to the S-locus. This finding is discussed with reference to recent reports that unilateral incompatibility is controlled, in peruvianum styles, by a number of different dominant genes and it is concluded that these dominant genes, the S-locus of self-incompatibility and the gametophytic factor regulating the unilateral reaction in esculentum pollen belong to the same linkage group. The strong sterility barriers which prevent practically all backcrosses between the hybrid and the parental species were shown to be independent of the factors regulating stylar incompatibility. L. peruvianum is heterozygous for the sterility genes which prevent fertilization or embryo formation when the interspecific hybrid is crossed, as pistillate parent, to different accessions of L. peruvianum. One peruvianum stock was found which, as a pollinator, was highly cross-fertile with the hybrids. The presence of a concentric endoplasmic reticulum in inhibited pollen tubes was observed to be a constant feature of both the self- and the unilateral incompatibility reactions and was interpreted as an indication that incompatibility might lead to a general cessation of protein synthesis. Although incompatible tubes very much resemble, in this respect, the pollen tubes cultured in vitro, it seems probable, on theoretical grounds, that the inhibition of pollen tubes in incompatible styles does not result from an absence of growth promoting substances but from the presence of a metabolic inhibitor.
    Type of Medium: Electronic Resource
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