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Size selection by continuous-flow centrifugation was used to prepare synchronous cultures of Bacillus subtilis. A selection of 5–10% was required to yield cultures which gave synchrony indices of about 045. Respiration rates [nmol O2 min-1 (ml culture)-1] of cells growing in the presence of glucose rose discontinuously as two steps in each cell cycle with the mid-points of the rises centred at 037 and 087 of a cycle. An asynchronous control culture exhibited no such periodicities. Two inhibitors of energy conservation—N,N′-dicyclohexylcarbodi-imide and 4-chloro-7-nitrobenzofurazan—inhibited respiration maximally at 00 and 06 of a cycle whilst an inhibitor of respiration—2-heptyl-4-hydroxyquinoline-N-oxide—exerted maximal effect only at 04 of a cycle. These results are compared with other temporal events which have been reported to occur during the cell cycle of B. subtilis.

Differential Effects of Inhibitors on Respiratory Activity of Synchronous Cultures of Bacillus subtilis Prepared by Continuous-flow Centrifugation