In order to facilitate the preparation of paeoniflorin (PF) and albiflorin (AF), two chief bioactive constituents in Paeonia lactiflora Pall (PL), induction and culture of callus from PL were studied. With a modified woody plant medium supplemented with 0.5 mg.L-1 6-benzylaminopurine, 1.0 mg.L-1 naphthylacetic acid, 0.1 mg.L-1 thidiazuron and 30 g.L-1 sucrose, callus was induced from four kinds of explants: leaf, stems, petiole, and root. The potency to form callus varies between different explants and leaf explants exhibits the highest capacity (100%). On the other hand, root-derived callus (R-callus) produces the highest level of total amount of PF and AF, 31.8 mg.g(-1) dry mass, which is higher than the corresponding level in the root of field cultivated PL. Furthermore, the time needed is only 40 days, remarkably shorter than the cultivation time of PL, about 4-5 years. Higher accumulation levels of PF and AF with shorter production time indicate that callus culture of PL is a promising powerful tool for production of PF and AF in the future. (C) 2014 The Chemical Industry and Engineering Society of China, and Chemical Industry Press. All rights reserved.