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Sequential cold-sensitive mutations in Aspergillus fumigatus.

O Nielsen, K F Gregory

    Canadian Journal of Microbiology
    |February 1, 1978
    PubMed
    Summary
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    Researchers developed cold-sensitive mutants of the thermotolerant fungus Aspergillus fumigatus. These mutants are unable to grow at 37°C, aiding in the study of fungal growth regulation.

    Area of Science:

    • Microbiology
    • Mycology
    • Fungal Genetics

    Background:

    • Thermotolerant fungi, such as Aspergillus fumigatus, play significant roles in various environments.
    • Understanding the genetic basis of fungal temperature sensitivity is crucial for controlling their growth and applications.
    • Previous studies have explored temperature adaptation in fungi, but targeted isolation of specific thermosensitive mutants remains challenging.

    Purpose of the Study:

    • To isolate and characterize cold-sensitive mutants of Aspergillus fumigatus I-21 (ATCC 32722).
    • To develop a method for obtaining mutants unable to grow at a specific non-permissive temperature (37°C).
    • To investigate the potential for sequential mutations to enhance cold sensitivity.

    Main Methods:

    • Gamma irradiation of Aspergillus fumigatus conidia to induce mutations.

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  • Enrichment of cold-sensitive mutants through alternating incubations at non-permissive temperatures and filtration.
  • Isolation of mutants with progressively lower growth temperatures.
  • Characterization of growth temperatures (minimum, optimum, maximum) for parent and mutant strains.
  • Assessment of mutant growth rates at elevated temperatures and reversion analysis.

    • Main Results:

    • The wild-type parent fungus has approximate growth temperatures of 12°C (min), 40°C (opt), and 50°C (max).
    • Direct isolation of mutants unable to grow at 37°C from the wild type was unsuccessful.
    • A mutant unable to grow at 25°C was isolated, and subsequent mutations increased cold sensitivity by 3-5°C increments.
    • Mutants completely unable to grow at 37°C were obtained after five sequential mutations.
    • All isolated mutants exhibited growth rates comparable to the wild type at temperatures of 45°C and higher.
    • Revertants capable of growing at the non-permissive and lower temperatures were produced by each mutant.

    Conclusions:

    • Sequential mutagenesis is an effective strategy for isolating Aspergillus fumigatus mutants with specific temperature sensitivities.
    • The developed method allows for the isolation of mutants with defined growth limitations, useful for genetic studies.
    • The observed reversion indicates genetic instability and provides tools for further analysis of temperature-regulated growth pathways in fungi.