Tomato systemin is a bioactive peptide that regulates the systemic activation of wound-responsive genes. It is released from its 200 amino acid precursor called prosystemin. Initial tissue-localization and hormone-induced expression assays indicated that the tomato prosystemin gene (SlPS) accumulates mainly in floral tissues and in response to exogenous abscisic acid and methyl jasmonate (MeJA) treatments, respectively. Later, the promoter regions of the PS gene in tomato (Solanum lycopersicum L. cv. Castlemart), pepper (Capsicum annuum) and potato (Solanum tuberosum) were isolated and an in silico analysis of the SlPS promoter revealed an over-representation of stress- and MeJA-responsive motifs. A subsequent 5′ deletion analysis of the SlPS promoter fused to the β-glucuronidase reporter (GUS) gene showed that the –221 to +40 bp proximal SlPS promoter region was sufficient to direct the stigma, vascular bundle-specific and MeJA-responsive expression of GUS in transgenic tobacco plants. Important vascular-tissue-specific, light- and MeJA-responsive cis-elements were also present in this region. These findings provide relevant information regarding the transcriptional regulation mechanisms of the SlPS promoter operating in transgenic tobacco plants. They also suggest that its tissue-specificity and inducible nature could have wide applicability in plant biotechnology.