Apparent antimutagenic effect of ultraviolet irradiation

Strain SL3367 is a S. typhimurium LT2 hisG46 stock which spontaneously reverts to His+ at a high frequency. Plates of defined medium with 1% (v/v) nutrient broth inoculated with ca. 10(8) washed SL3367 cells were incubated, untreated or after UV irradiation. After 2 days at 37 degrees C, an average of 165 His+ colonies were obtained per control plate but significantly fewer, 105 His+ colonies, on plates irradiated at a fluence of 7 J/m2. The dry weight of bacteria in washings from plates incubated 14 h (by which time growth of His- cells had ceased) was the same for irradiated and non-irradiated plates but the yield of colony-forming units from irradiated plates was less than from control plates, by about the same factor as the reduction in yield of His+ colonies caused by the same fluence. Washings from incubated irradiated plates, but not those from control plates, contained long filaments as well as bacteria of normal size; on transfer to nutrient-agar slide cultures cells of normal size grew into microcolonies but filaments did not grow. The reduced plateau yield of viable His- cells caused by consumption of much of the growth-limiting supply of histidine by irradiated cells growing into non-viable filaments reduces the number of auxotrophic bacteria at risk for spontaneous reversion and so accounts for the apparent antimutagenic effect of UV irradiation. This effect was partly reversed by the presence of D,L-pantoyl lactone in the selection medium, and was also observed for yield of Trp+ colonies from trpE8 cultures with a high spontaneous reversion rate. Treatments not inducing cell filamentation did not result in the depression of spontaneous revertants and were detected as being mutagenic. The apparent antimutagenic effect may be expected for reversion of any auxotroph, unless masked by induced revertants and is particularly apparent in an auxotroph which reverts spontaneously at high frequency.