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  • 1
    Electronic Resource
    Electronic Resource
    Protoplast technology in relation to crop plants: Progress and problems (1977)
    Springer
    Euphytica 26 (1977), S. 343-360 
    Details
    ISSN: 1573-5060
    Keywords: Protoplasts ; somatic hybridisation ; culture of cereal protoplasts ; regeneration ; fusion ; heterokaryon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The ready availability of isolated plant protoplasts, and the ability of these naked plant cells to fuse together, has greatly stimulated interest in the production of plant somatic hybrids. This new protoplast technology may enable interspecies hybrids to be obtained which are otherwise not possible. Many such interspecies hybrids are required amongst the crop plants, but application of this new technology to these species in presently prevented by our inability to culture cells of many of these species in vitro, and to regenerate plants reproducibly from cultured tissue.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00026995
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    The interaction betweenE. coli spheroplasts and plant protoplasts: A proposed procedure to deliver foreign genes into plant cells (1986)
    Springer
    Protoplasma 130 (1986), S. 153-161 
    Details
    ISSN: 1615-6102
    Keywords: DNA vector delivery ; Fusogens ; Protoplasts ; Spheroplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary E. coli spheroplasts can be used to deliver DNA vectors into plant protoplasts. The use of fluorescent dyes showed that 25–100% of the protoplast population was associated with 1–9 spheroplasts following incubation with several fusogens. Electron microscopy demonstrated spheroplasts attached to protoplasts via a plasma membrane protrusion after high pH/Ca2+ treatment, but PEG-high pH/Ca2+ promoted endocytosis of spheroplasts into a plasma membrane bounded vesicle. Ultrastructural profiles showed that fusion between spheroplasts and protoplasts did not occur. Immunofluorescence studies detectedE. coli antigens associated with tobacco protoplasts, and after fusogen treatment the antigens were dispersed within the peripheral cytoplasm. The elimination of residual contaminatingE. coli cells from protoplasts was achieved by lysozyme and antibiotic treatment, thus allowing DNA vector assessment in axenic culture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01276597
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  • 3
    Electronic Resource
    Electronic Resource
    Flow cytometric quantification of electroporation-mediated uptake of macromolecules into plant protoplasts (1995)
    Springer
    Protoplasma 186 (1995), S. 50-56 
    Details
    ISSN: 1615-6102
    Keywords: Electroporation ; Flow cytometry ; Macromolecules ; Protoplasts ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Flow cytometry was used to provide a rapid and accurate assessment of electroporation-induced uptake of macromolecules into plant protoplasts. Rice protoplasts were electroporated in the presence of fluorescein isothiocyanate-conjugated dextran (FITC-dextran). After washing, the protoplasts were resuspended in a solution containing propidium iodide which intercalates with DNA, but which is excluded by an intact plasma membrane. Electroporation in the presence of FITC-dextran gave rise to populations of protoplasts that fluoresced green or yellow due to the presence of non-conjugated FITC. Non-viable protoplasts fluoresced red because of their inability to exclude propidium iodide molecules. Flow cytometry was used to resolve and quantify these protoplast populations and thus identify optimal conditions for macromolecule uptake. A direct relationship was observed between FITC-dextran uptake and transient gene expression following plasmid uptake. Thus, simultaneous electroporation of protoplasts with foreign DNA and FITC-dextran followed by fluorescence activated cell sorting may permit partial selection of transformed cells and so reduce the need for a selectable marker.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01276935
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