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  • 1
    Digitale Medien
    Digitale Medien
    Springer
    Theoretical and applied genetics 79 (1990), S. 625-631 
    ISSN: 1432-2242
    Schlagwort(e): Microprojectile bombardment ; Bialaphos ; bar gene ; Maize ; Transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Stable transformed Black Mexican Sweet (BMS) maize callus was recovered from suspension culture cells bombarded with plasmid DNA that conferred resistance to the herbicide bialaphos. Suspension culture cells were bombarded with a mixture of two plasmids. One plasmid contained a selectable marker gene, bar, which encoded phosphinothricin acetyl transferase (PAT), and the other plasmid encoded a screenable marker for β-glucuronidase (GUS). Bombarded cells were selected on medium containing the herbicide bialaphos, which is cleaved in plant cells to yield phosphinothricin (PPT), an inhibitor of glutamine synthetase. The bialaphos-resistant callus contained the bar gene and expressed PAT as assayed by PPT inactivation. Transformants that expressed high levels of PAT grew more rapidly on increasing concentrations of bialaphos than transformants expressing low levels of PAT. Fifty percent of the bialaphos-resistant transformants tested (8 of 16) expressed the nonselected gene encoding GUS.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    ISSN: 1615-6102
    Schlagwort(e): Freeze-fracture ; Isolated rye protoplasts ; Osmotic contraction ; Plasma membrane-derived vesicles ; Ultrastructure
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Following osmotic contraction of isolated rye protoplast (Secale cereale L. cv. Puma) that results in nearly a 50% reduction in volume, the plasma membrane was smooth, with no folding or pleating. Instead, deletion of plasma membrane occurred and numerous cytoplasmic vesicles were observed. As a result, the area of the plasma membrane was reduced by approximately 40%. Thin sections revealed that the cytoplasmic vesicles were membrane bound and not merely voids in the cytoplasm. High resolution video microscopy revealed the extent of vesiculation showing large clusters of cytoplasmic vesicles following osmotic contraction. Labeling the plasma membrane with fluorescein-Con-A prior to hypertonic contraction suggested that the cytoplasmic vesicles were derived from the plasma membrane. Freeze-fracture particle density on both the protoplasmic (PFp) and exoplasmic face (EFp) of the plasma membrane remained unchanged following contraction, which is consistent with a unit-membrane deletion into cytoplasmic vesicles. Upon partial re-expansion of the protoplasts, thin sections showed that the vesicles remained in the cytoplasm. These results using osmotic manipulation confirm earlier observations of isolated protoplasts at the light microscope level. Upon contraction plasma membrane is deleted into cytoplasmic vesicles, which are not readily reincorporated into the plasma membrane upon expansion. Lysis occurs before the original volume and surface area are regained.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 3
    ISSN: 1615-6102
    Schlagwort(e): Cold acclimation ; Exocytotic extrusions ; Freeze-fracture ; Isolated rye protoplasts ; Lipid bodies ; Osmotic contraction ; Plasma membrane ; Ultrastructure
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary Osmotic contraction of protoplasts isolated from cold acclimated leaves ofSecale cereale L. cv. Puma results in the formation of exocytotic extrusions of the plasma membrane. Numerous knobs or polyps were observed on the surface of the protoplasts with scanning electron microscopy. In thin sections, the extrusions were bounded by the plasma membrane with a densely osmiophilic interior. Cross-fracturing of the extrusions revealed aparticulate bodies within, a further indication that the interior of the extrusions was predominantly lipid material. Freeze-fracture of the plasma membrane suggests a possible source of this lipid material. Following osmotic contraction, the particle density on the plasma membrane protoplasmic face (PFp) increased, being reflected in both a substantial increase in paracrystalline arrays and an increase in the particle density in non-crystalline regions. This increase in particle density indicates that lipid material is preferentially lost from the plasma membrane during contraction. The density on the exoplasmic face (EFp) did not change. Together, these findings suggest that during hypertonic contraction of acclimated protoplasts, lipid material is preferentially subducted from the plasma membrane and sequestered into lipid bodies (the osmiophilic regions). The formation of lipid bodies and extrusions was readily reversible. Following osmotic expansion of acclimated protoplasts, the extrusions were retracted back into the plane of the plasma membrane.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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