Abstract: The rooting and physiological biochemistry metabolize changes were studied during rooting of the hardwood of Thompson seedless grape (Vitis vinifera L.) by treating of ABT and CaCN2 with water control. The results indicated that the cuttings by treating of ABT rooted earlier, rooting date and the number of roots more than other treatments, and the roots grew healthily. The cuttings by treating of CaCN2 buded earlier, and buding date and the length of branchs were more than others.Root rates and bud rates by treating of CaCN2 firstly , then by treating of ABT were between ABT and CaCN2, respectively, but there were significant difference with water control, and no significance ABT and CaCN2, respectively. Starch could change to soluble sugar easily by treating of ABT singly or both ABT and CaCN2, so promoting rooting. The activity of indoleacetic acid oxidase ( IAAO ) , polyphenol oxidase (PPO ) , as w ell as, the relationship between enzyme activities and rooting capacities of Thompson seedless grape cuttings was studied in this paper. There were the more relationship between rooting capacities and enzyme activities, which changed with different rooting time. During callus formatting, higher IAAO activities could release IAA in grape, benefiting inducing roots, but during root elongating, lower IAAO activities benefited to protect IAA been released, promoting roots growing. During the course of adventitious root formation, PPO activities increase quickly, promoting rooting.