A unidirectional promoter can be transformed into a bidirectional module by artificial methods. Here we report the bidirectionalization of the methyl jasmonate (MeJA)-inducible PtDrl02 promoter derived from poplar [(Populus tomentosa × P. bolleana) × P. tomentosa] in planta. Construction of the bidirectional PtDrl02 promoter (designated as mPtDrl02) was rapidly achieved by introducing a minimal 35S promoter in the opposite orientation to the 5' end of PtDrl02. β-Glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes were also interchangeably linked to the 3'-and 5'-ends of mPtDrl02 to produce GFP/mPtDrl02/GUS and GUS/mPtDrl02/GFP vectors, respectively. Using the Agrobacterium-mediated transient expression approach, we demonstrated that the mPtDrl02 module was able to drive gene (GUS and GFP) expression in both orientations simultaneously. Furthermore, the cooperative and concurrent activity from both directions of the mPtDrl02 module was demonstrated following MeJA induction. To our knowledge, this is the first report of an artificial MeJA-responsive bidirectional promoter in perennial plants.