Gardenia (Gardenia jasminoides) is a woody medicinal plant with geniposide, chlorogenic acid and crocin-1 as its bioactive compounds. To establish an experimental system for gene transformation to obtain high-content bioactive compounds, we studied the effects of different plant regulators and cultural modes on the callus induction and shoot regeneration from explants of pericarps, placentas with seeds, and seeds of gardenia. The optimal medium for callus induction from pericarp and seed was MS+0.5 mg·L^–12,4-D+0.25 mg·L^–16-BA. However, for callus induction from placentas with seeds, the optimal medium was MS+1.0 mg·L^–12,4-D+1.0 mg·L^–16-BA. The rates of callus induction were 83.3%, 88.5% and 78.1% from explants of pericarp, seed, and placenta with seeds, respectively. Buds regenerated from seed callus by liquid culture but not from the other 2 kinds of calli. TDZ (N-phenyl-N′-1,2,3-thidiazol-5ylurea), a phenylurea compound with auxin and cytokinin activities, stimulated bud differentiation. The optimal medium for bud differentiation was MS+0.05 mg·L^–1NAA+0.10 mg·L^–1TDZ with highest differentiation rate of 8.75%. We established a plant tissue culture system with gardenia seeds as explants, which can serve as a technical platform for gene transformation.