Abstract: In order to study the effect of plant growth regulators on the rooting of Vitex quinata, the cuttings of semilignified Vitex quinata were treated with different kinds, concentrations and soaking time of plant growth regulators to study the rooting and physiological and biochemical changes of cuttings. The results showed that there were significant differences in rooting rate, average root length, longest root length, average root number, average root diameter and longest root system diameter among different plant growth regulators. Through the comprehensive evaluation of membership function method, the membership function value of the treatment with NAA concentration of 200 mg/L and soaking time of 2 h was the highest, reaching 0.88. In the process of rooting, the content of soluble sugar and soluble protein showed a trend of ‘decline-rise-decline’. The activities of SOD, POD and PPO in the treatment group were higher than those in CK treatment. In the root primordial induction stage, the activities of SOD and POD increased, while in the adventitious root formation stage, the activities of SOD and PPO continued to increase and the activities of POD decreased. In the adventitious root elongation stage, the activities of POD increased and the activities of SOD and PPO decreased. NAA concentration of 200 mg/L and soaking time of 2 h were the most suitable treatments for cutting rooting of Vitex quinata, which can change the nutrient content of cuttings, improve the enzyme activity during rooting and promote the growth of rooting.