Identification of different sites of expression for spo loci by transformation of Bacillus subtilis

Abstract

Asporogenous mutants of Bacillus subtilis were rendered capable of forming heat-resistant spores by transformation with wild-type (spo+) DNA at, or near, the start of sporulation. For several mutants up to about 50% of the colonies derived from heat-resistant spores, formed as a result of the transformation, remained genetically asporogenous (spo). This was thought to indicate that the genome of the mother cell, but not that of the forespore, was transformed to spo+ and that correct expression of the spo locus in the mother cell was sufficient for spore formation. At the end of the process the mother cell was destroyed, leaving a mature heat-resistant spore that was genetically asporogenous. It is concluded that the loci spoIIID, spIVA, spoVB and spoVE are expressed in the mother cell. For one mutant more than 99% of the colonies derived from heat-resistant spores were genetically spo+. It is concluded that the locus involved, spoVA, had to be expressed in the forespore. Thus different sporulation-specific loci are expressed in the mother cell and in the forespore. The loci expressed in the mother cell are expressed in one cell type so that another cell type, the forespore, can develop into a heat-resistant spore. Other unselected donor markers could be introduced into the recipient during transformation provided high concentrations of DNA were used. The frequency of congression was the same for spo survivors as for spo+ survivors. This implies that there was no correlation between the DNA strand into which the selected spo+ and the unselected donor markers integrated.