In order to explore the rejuvenation effect based on three levels which included the induction rate of multiple shoots and the rooting rate, morphological indexes and physiological indexes, we used Armeniaca sibirica as trial material and selected the tissue culture seedling of different positions, ages and subculture times. The results showed as follows: (1) the indexes of every rejuvenation effect showed the same variation rule in tissue culture seedling. When the sampling position of A. sibirica explants decreased which was from the crown of uppermiddlelower, the induction rate of multiple shoots and the rooting rate of tissue culture seedling were keeping rising in turn. The difference among positions was significant. The value of plant height, leaf length, leaf width, root number and root length also increased along with the decline of the explant sampling position. Only the difference of root number and root length were significant between positions. But the SOD activity, POD activity and MDA content of leaves decreased gradually, and those in leaves of tissue culture seedling were always lower than that in seed tree leaves. (2) With the decreasing of seed tree age which was from 10a-4a-1a, the above indexes values of tissue culture seedling all raised in turn. The rooting rate, root number root length of tissue culture seedling and the induction rate of multiple shoots which was from 1a were higher significantly than the corresponding indexes of 10a and 4a. The values were 80%, 96.6%, 4.3 and 2.40 cm, respectively. (3) From primary culture to first and second subculture, the induction rate of multiple shoots, plant height, leaf length and leaf width of various tissue culture seedlings increased constantly with the increasing of subculture times and the second subculture had larger increasing. The SOD activity, MDA content and POD activity of leaves appeared downward trend in tissue culture seedling, but still behaved that the tissue culture seedling of second subculture had the better rejuvenation effect. In summary, with the moving down of explant sampling positions, the decreasing of sampling age in seed tree and the increasing of subculture times in tissue culture seedling, the induction rate of multiple shoots and the rooting rate, morphological indexes, physiological indexes of tissue culture seedling showed the consistent trends, and the rejuvenation effect of tissue culture was better.