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The aim of the study was to examine the effect of opposition and gender on knee kinematics and ground reaction force during landing from a volleyball block jump. Six female and six male university volleyball players performed two landing tasks 1) an unopposed and 2) an opposed volleyball block jump and landing. Knee kinematics were recorded by a 12 camera motion analysis system (120 Hz) and ground reaction force was recorded by a force platform (600 Hz) during landing. The results showed a significant effect for level of opposition in peak normalized GRF (p = .04), knee flexion at ground contact (p = .003), maximum knee flexion (p = .001) and range of motion of knee flexion (p = .003). There was a significant effect for gender in maximum knee flexion (p = .01), range of motion of knee flexion (p = .001), maximum knee valgus angle (p = .001) and range of motion of knee valgus (p = .001). The changes in landing biomechanics as a result of opposition suggest future research investigating landing mechanics should examine opposed exercises since opposition may significantly alter neuromuscular responses.