Detection of genotoxicants in Brassicales using endogenous DNA as a surrogate target and adducts determined by 32P-postlabelling as an experimental end point

Abstract

Some plants use electrophilic metabolites as a defence against biological enemies. Some of them may react with DNA. We devised a new model to test this hypothesis. Plant tissue was homogenised. After incubation of the homogenate at 37°C for varying periods, the plant DNA was analysed for the presence of adducts using the (32)P-postlabelling technique. Adducts were detected with all Brassicales studied. Broccoli was investigated in detail. Adducts were absent in DNA isolated immediately after homogenisation of the plant. Subsequently, five characteristic adduct spots were formed in the homogenate, the maximum being reached after nearly 4 h. Adduct formation was low when broccoli was steamed before homogenisation, but was re-established when myrosinase was added to the homogenate, indicating that the active constituents were glucosinolates. Broccoli juice was mutagenic to Salmonella typhimurium, forming the same adduct spots in these target cells as in plant homogenate, but the relative intensity of the individual spots varied between both models. The patterns of adduct spots formed in homogenates of 15 other Brassicales species and tissues were similar to those detected with broccoli florets heads. However, the relative intensities of the spots varied. Sporadically, some spots were missing or additional spots appeared. These results, therefore, suggest that several different glucosinolates contribute to the adduct formation.