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Intracellular centrifugal separation of organelles in Phycomyces

M Zalokar

    The Journal of Cell Biology
    |May 1, 1969
    PubMed

    Insights

    Centrifugation of living Phycomyces blakesleeanus sporangiophores separated cell components into distinct layers. These isolated organelles remained viable, enabling biochemical studies and demonstrating functional potential, such as nuclei inducing color formation.

    Area of Science:

    • Cell Biology
    • Biochemistry
    • Mycology

    Background:

    • Phycomyces blakesleeanus is a model organism for studying cellular processes.
    • Understanding the spatial organization and viability of cell organelles is crucial for biochemical analysis.

    Purpose of the Study:

    • To isolate and characterize cellular components of Phycomyces blakesleeanus using high-speed centrifugation.
    • To assess the viability and functional capacity of sedimented organelles for biochemical studies.

    Main Methods:

    • Living sporangiophores of Phycomyces blakesleeanus were subjected to ultracentrifugation (35,000 rpm).
    • Sedimented layers were analyzed using electron microscopy and cytochemical techniques.
    • Organelle viability was tested by injecting nuclei into a host sporangiophore and observing induced color formation.

    Main Results:

    • Centrifugation yielded distinct layers of cellular components including polyphosphates, glycogen, ribosomes, mitochondria, nuclei, and endoplasmic reticulum.
    • Isolated nuclei from a lycopene-producing mutant induced color formation when injected into an albino host.
    • Microspectrophotometry revealed cytochromes and flavines in mitochondria and cytochromes in nuclei.

    Conclusions:

    • Cell organelles isolated by centrifugation of living sporangiophores remain viable and suitable for biochemical investigations.
    • The functional integrity of isolated organelles, like nuclei, can be confirmed through biological assays.
    • This method provides a valuable approach for studying organelle function and composition in Phycomyces blakesleeanus.

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