A reliable method for measuring the spore DNA content, based on radioactive DNA labelling, spore germination in absence of DNA replication and diphenylamine assay, was developed. The accuracy of the method, within 10-15%, is adequate for determining the number of chromosomes per spore, provided that the genome size is known. B subtilis spores were shown to be invariably monogenomic, while those of larger bacilli Bacillus megaterium, Bacillus cereus and Bacillus thuringiensis, often, if not invariably, contain two genomes. Attempts to modify the spore DNA content of B subtilis by altering the richness of the sporulation medium, the sporulation conditions (liquid or solid medium), or by mutation, were apparently unsuccessful. An increase of spore size with medium richness, not accompanied by an increase in DNA content, was observed. The implication of the apparently species-specific spore ploidy and the influence of the sporulation conditions on spore size and shape are discussed.

Bacillus↗

PubMedDate not supplied

Morphological and cell wall alterations in thermosensitive DNA mutants of Bacillus subtilis.

Incubation of thermosensitive dna mutants of Bacillus subtilis at the nonpermissive temperature results, at the cellular level, in the appearance of swellings. The study of one particular type of swelling, named "terminal balloon", reveals that the occurrence of the latter was correlated with completion of rounds of DNA replication. Morphological and autoradiographic observations reveal that (a) cell wall consists of two layers, (b) the outer layer splits at a fixed distance from the cell pole and allows the formation of a balloon contained within a single wall layer and (c) the bulk of active synthesis of cell wall is localized in the balloon area leading to the formation of a near spherical monolayer. Implications of these results for cell wall morphogenesis are discussed.

Bacillus subtilis↗