The potential use of a new chemical-inducible gene expression system in Arabidopsis thaliana has been examined. The system is based on the maize In2-2 promoter which is activated by benzenesulfonamide herbicide safeners. Plants transformed with the beta-glucuronidase (gus) reporter gene under the control of the In2-2 promoter were grown in the presence of different safeners and the induced GUS activity pattern was studied histochemically. In the absence of safeners, the In2-2 promoter was not active. Application of different safeners induced distinct gus expression patterns, including expression in the root, hydathodes, and the shoot apical meristem. Plants maintained continuously on inducing concentrations of the safeners were retarded in growth. The growth inhibition effects of the Sa5 safener could be overcome in a sulfonylurea-resistant background. In2-2 promoter activity could also be induced by the sulfonylurea herbicide chlorsulfuron. In the sulfonylurea-resistant background, which derives from herbicide-resistant acetolactate synthase activity, induction of the In2-2 promoter by chlorsulfuron was lower. Furthermore, branched-chain amino acids, known to inhibit acetolactate synthase activity, also induced In2-2 promoter activity. Our data suggest a strong correlation between In2-2 expression and inhibition of the acetolactate synthase activity.