Mutagenesis in multinucleate cells: the effects of N-methyl-N'-nitro-N-nitrosoguanidine on Phycomyces spores

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M I Roncero, C Zabala, E Cerdá-Olmedo

    Mutation Research
    |February 1, 1984
    PubMed

    Insights

    This study introduces a method using N-methyl-N'-nitro-N-nitrosoguanidine to create single-nucleus fungal cells, enabling the study of recessive mutations in Phycomyces. This technique improves the isolation of desired genetic mutations.

    Area of Science:

    • * Mycology
    • * Molecular Biology
    • * Genetics

    Background:

    • * Multinucleate cells, like Phycomyces spores, present challenges for isolating recessive mutants due to multiple gene copies.
    • * Traditional mutagenesis methods are hindered by the presence of multiple nuclei, masking recessive traits.

    Purpose of the Study:

    • * To develop a method for isolating recessive mutants in multinucleate fungal cells.
    • * To optimize mutagenesis conditions for Phycomyces using N-methyl-N itro-N-nitrosoguanidine.
    • * To enable the study of recessive mutations by achieving functional uninucleate cells.

    Main Methods:

    • * Employing N-methyl-N itro-N-nitrosoguanidine (nitrosoguanidine) for nuclear killing in Phycomyces spores.
    • * Selecting for survivors with a single functional nucleus after nitrosoguanidine treatment.
    • * Utilizing heterokaryon spores and recessive markers for positive selection of uninucleate cells.

    Main Results:

    • * Nitrosoguanidine effectively reduces nuclei in some Phycomyces cells, allowing recessive mutation expression.
    • * Approximately 35% of nitrosoguanidine-treated survivors achieve functional uninucleate status.
    • * Optimal nitrosoguanidine mutagenesis conditions in Phycomyces differ from bacteria and yeast, with less emphasis on buffer pH.

    Conclusions:

    • * Nuclear killing with nitrosoguanidine is a viable strategy for isolating recessive mutants in multinucleate fungi.
    • * The developed method facilitates the study of recessive mutations by enabling the isolation of functionally uninucleate cells.
    • * Understanding mutagenesis kinetics (survival and mutation induction) is crucial for optimizing genetic studies in Phycomyces.

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