Jatropha curcas is a second generation biofuel crop that is also an important biological resource. Meeting increasing biofuel demand requires wider cultivation of specialized varieties, highlighting the need for massive culture techniques such as in vitro micropropagation. Key stages in plant tissue culture processes, such as adventitious shoot induction, can be optimized to increase micropropagation protocol efficiency. Response surface methodology, a statistical tool for this purpose, was applied to optimize in vitro induction of adventitious shoots in the J. curcas micropropagation process. A preliminary assay was done in which 6-(γ, γ-dimethylallylamino) purine and indolacetic acid were selected as the factors to optimize. A 2 2 central composite design with four central points was then applied to fit it to a second grade model and a rotatable design applied to produce the final model and surface graphics. Focused on maximum point, the surface model design identified maximum adventitious shoot induction to be 21 shoots per explant in Murashige and Skoog medium at 75% ionic strength and containing 83.3 µM 6-(γ, γ-dimethylallylamino) purine and 1.3 µM indolacetic acid. This is the first report in which response surface methodology was found to be an effective statistical technique for optimizing adventitious shoot induction in Jatropha curcas.