The paper aims to establish a technical system for the cultivation of paeonol in P. ostii var. lishizhenii callus,and to provide a technical basis for cell engineering pharmacy.The mature and plump seeds of P. ostii var. lishizhenii were peeled and soaked in 70% ethanol(V/V) for 2.0 min and then soaked in 1.0% sodium dichloroisocyanurate solution for 20 minutes.The seed decontamination rate was 97.78%.Place sterilized on MS medium and then add 6-BA 1.5 mg/L and 2,4-D 2.0 mg/L.The callus induced was 90.00%.Optimized medium for cell culture was the modified WPM with 6-BA 1.8 mg/L,NAA 0.6 mg/L,sucrose 30 g/L,hydrolyzed casein 0.4 g/L,agar 5 g/L and pH 7.3.In the conditions,cell proliferation ratio was 3.53 and the browning rate was 11.78% on the 25th day.Based on the natural product paeonol content and callus growth curve,paeonol content reached the highest when cultured for 25 days,which was the best harvest time.