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Women sustain more anterior cruciate ligament (ACL) ruptures than men, and this gender disparity is apparent from pubertal onset. Although the hamstring muscles play a vital role in ACL protection during landing by restraining anterior tibial motion relative to the femur, it is unknown whether hamstring strength affects landing biomechanics during a functional movement. This study aimed to determine whether pubescent girls with lower hamstring strength displayed different lower limb biomechanics when landing from a leap compared with girls with higher hamstring strength.Thirty-three healthy girls, age 10–13 yr, in Tanner stage II (pubertal onset) and 4–6 months from their peak height velocity were recruited. The concentric and the eccentric isokinetic strength of the hamstring and quadriceps muscles were assessed. On the basis of peak concentric hamstrings torque, participants were divided into a lower (peak torque < 45 N·m) and higher (peak torque > 60 N·m) strength group. Participants performed a functional landing movement, during which ground reaction forces (1000 Hz), lower limb electromyography (1000 Hz), and kinematic data (100 Hz) were collected.Girls with lower hamstring strength displayed significantly (P < 0.05) greater knee abduction alignment, reduced hip abduction moments, and greater ACL loading at the time of the peak anteroposterior ground reaction forces compared with their stronger counterparts.Girls with reduced hamstring strength appear to have a decreased capacity to control lower limb frontal plane alignment. This reduced capacity appears to contribute to increased ACL loading and, in turn, increased potential for injury.