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  • Freeze substitution  (2)
  • 2010-2014
  • 1990-1994  (2)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 163 (1991), S. 199-202 
    ISSN: 1615-6102
    Keywords: Actin ; Freeze substitution ; Fungi ; Hyphal tip ; Immunocytochemistry ; Magnaporthe grisea
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have successfully localized fungal actin for the first time using immuno-electron microscopy and hyphal tips of the rice blast pathogenMagnaporthe grisea. Following ultrarapid freezing, samples were processed in a novel substitution fluid of 10% acrolein in anhydrous ethanol and embedded in LR White resin. A monoclonal anti-actin antibody, previously shown to recognizeM. grisea actin, bound specifically to filasomes concentrated in the peripheral cytoplasm of subapical regions, and to the core-region of the Spitzenkörper.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Protoplasma 168 (1992), S. 20-26 
    ISSN: 1615-6102
    Keywords: Appressoria ; Freeze substitution ; Immunofluorescence ; Plant disease ; Phalloidin ; Pyricularia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The penetration peg is the structure used byMagnaporthe grisea to pierce the surface of rice leaves or very hard nonbiodegradable substrates. Penetration pegs produced by appressoria in vitro were examined by electron microscopy and immunofluorescence microscopy using various fluorophore labeled anti-actins. Freeze-substitution preparation of appressoria at early stages of substrate penetration showed that peg cytoplasm consisted primarily of a zone of exclusion, excluding even ribosomes, and was continuous with a similar region in the appressorium. Apical vesicles were, however, observed in short, presumably elongating pegs. Immunofluorescence microscopy was used to demonstrate binding of a monoclonal anti-actin to penetration peg cytoplasm, following “permeabilization” of appressoria by means of a brief sonication. Occasional filaments and ca. 300 nm diameter plaques were labeled in appressorial cytoplasm. Western blot analysis of germ tube extracts showed that the monoclonal probe bound predominantly to a single band with a molecular weight similar to that of rabbit muscle actin. Preincubation of the antibody with actin virtually eliminated peg labeling. We conclude that the penetration peg contains actin which may play a role in the formation of the zone of exclusion.
    Type of Medium: Electronic Resource
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