The effects of different physical and chemical factors on callus growth and flavonoid production in Cyclocarya paliurus were studied.The results showed that(1) The optimal subculture times of callus growth was the eighth,the flavonoid content of the tenth times was the highest,the peak content was 2.02%;(2) The callus cultured in light for 12 h·d-1 were suitable for the callus growth and flavonoid accumulation;(3) The suitable pH value of medium for callus growth were 5.8,for flavonoid production was 6.0;(4) 3%sucrose and 5 mg·L-1 E material was optimal for callus growth and flavonoid formation;(5) The suitable material for callus subculture is from the stems in the middle of May,and for flavonoid accumulation is in the middle of July.