Thymineless death in Escherichia coli in various assay systems: viability determined in liquid medium

Thymineless death has been studied in four different Thy(-) strains of Escherichia coli by using various assay methods including conventional plating techniques as well as one performed entirely in liquid medium. Plating on L agar resulted in a greater loss in viability than the other assay methods, but this extrasensitivity of starved cells to L-agar plating quickly disappeared upon readdition of thymine to the starved cultures. This indicated that cellular damage responsible for the additional killing on L agar is reversible. The results obtained by three other assay methods, the liquid assay, plating on nutrient agar, or plating on tris(hydroxymethyl)aminomethane-minimal agar, did not differ significantly from each other with all strains tested except strain JG 151. In this strain thymineless death was much faster when assayed in the liquid system than by plating. It is suggested that thymineless death detected on nutrient or minimal agar is not a result of plating, but that the lethal event actually occurs during the period of thymine starvation.