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Cell wall and morphological changes induced by temperature shift in Bacillus subtilis cell wall mutants.

M A Shiflett, D Brooks, F E Young

    Journal of Bacteriology
    |November 1, 1977
    PubMed
    Summary

    Bacillus subtilis mutants exhibit altered cell wall composition and morphology at high temperatures. Revertants show varying teichoic acid levels, impacting bacteriophage sensitivity and cell shape.

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    Area of Science:

    • Microbiology
    • Cell Biology
    • Biochemistry

    Background:

    • Bacillus subtilis strains RUB1012 and RUB1013 display distinct phenotypes at 45°C, including impaired growth, altered cell morphology (clumps of spheres), reduced autolysis, and a low ratio of teichoic acid to peptidoglycan.
    • These temperature-sensitive cell wall defects suggest a role for teichoic acid in maintaining normal cell structure and function under stress.

    Purpose of the Study:

    • To investigate the genetic basis and phenotypic consequences of cell wall alterations in Bacillus subtilis at elevated temperatures.
    • To characterize the relationship between teichoic acid content, cell morphology, and bacteriophage susceptibility in wild-type and mutant strains.

    Main Methods:

    • Isolation and characterization of temperature-sensitive revertants of Bacillus subtilis RUB1012.

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  • Quantification of teichoic acid content in cell walls using phosphorus measurement.
  • Assessment of bacteriophage plating efficiency on different strains.
  • Microscopic analysis (scanning electron microscopy) of cell morphology during temperature shifts.

    • Main Results:

    • Revertants (RUB2032, RUB2012, RUB2042) regained the ability to grow at 45°C but exhibited varying proportions of teichoic acid to peptidoglycan, with significantly lower phosphorus content compared to the parent strain.
    • Bacteriophage plating efficiency correlated with the amount of glucosylated teichoic acid.
    • Shifting RUB2032 from 30°C to 45°C induced a morphological transition from rods to spheres, characterized by cell wall thickening and inhibited cell separation.
    • Shifting RUB2032 from 45°C to 30°C resulted in amorphous material accumulation preceding the rod morphology, with cells remaining clumped.

    Conclusions:

    • Multiple mutations affecting cell wall synthesis and composition are responsible for the observed temperature-sensitive phenotype in Bacillus subtilis RUB1012 and RUB1013.
    • Teichoic acid content and glucosylation are critical for maintaining cell shape, facilitating cell separation, and influencing susceptibility to bacteriophages.
    • The study elucidates the complex interplay between cell wall components, environmental stress, and bacterial morphology.