Efforts to elucidate the contributions by transcription factors to plant gene expression will require increasing knowledge of their specific in vivo regulatory associations. We are systematically investigating the role of individual TGA factors in the transcriptional control of pathogenesis-related (PR) defense genes, whose expression is stimulated in leaves by salicylic acid (SA) through a stimulus pathway involving NPR1. We focused on PR-1 because its SA-induced expression in Arabidopsis is mediated by an as-1-type promoter cis element (LS7) that is recognized in vitro by TGA factors. We found that two NPR1-interacting TGA factors, TGA2 and TGA3, are the principal contributors to an LS7 binding activity of leaves that is enhanced by SA through NPR1. The relevance of these findings to PR-1 expression was investigated by the use of chromatin immunoprecipitation, which demonstrated that in vivo these TGA factors are strongly recruited in an SA- and NPR1-dependent manner to the LS7-containing PR-1 promoter. Significantly, the timing of promoter occupancy by these factors is linked to the SA-induced onset and sustained expression of PR-1. Because leaf transfection assays indicate that TGA3 activates transcription, as noted previously for TGA2, these two TGA factors are predicted to make positive contributions to the expression of this target gene. Thus, the findings presented here distinguish among different modes of regulation by these transcription factors and provide strong support for their direct role in the stimulus-activated expression of an endogenous defense gene.