The spider mite Tetranychus urticae Koch is emerging as a major problem in Jatropha curcas cultivation. The goal of this study was to investigate the photosynthetic responses of Jatropha to spider mite infestation. Leaf CO2 assimilation rate, stomatal conductance, transpiration, intracellular CO2 concentration, and instantaneous carboxylation efficiency significantly decreased in mite-infested leaves compared with controls. Lower water content and specific leaf area of the mite-infested leaves were positively related to symptoms of wrinkling and curling. Leaf electrolyte leakage remained unchanged in the mite-infested leaves, revealing no effect on leaf membrane integrity. Leaves exhibited reductions in soluble protein and soluble sugar in association with photosynthetic impairment. Although decreases in photochemical activity and chlorophyll fluorescence parameters suggested damage to the photosynthetic apparatus, although there were no measurable reductions in chlorophyll or carotenoid contents associated with photosynthetic apparatus impairment. The decrease in the leaf CO2 assimilation rate was partially attributed to stomatal and metabolic limitations in the mite-infested leaves.