Adventitious shoot regeneration from in vitro stem explants of Phellodendron amurense

An efficient in vitro plant regeneration system from stem explants was established in Phellodendron amurense. Factors influencing shoot regeneration from stems including culture medium type, combinations of plant growth regulators and carbon source in the medium were investigated. Adventitious shoot regeneration was significantly influenced by the type of medium. Murashige and Skoog medium (MS) was the best for promoting shoot regeneration, followed by Gamborg medium (B5) and woody plant medium (WPM). The combination of 6-benzyladenine (BA) and naphthaleneacetic acid (NAA) produced better results for shoot regeneration. The optimum shoot regeneration frequency (74.5%) and number of shoots per explant (12.3) was achieved using MS medium supplemented with 29.7 M BA and 5.8 M NAA. High concentrations of BA and NAA in the medium inhibited shoot formation. Among the three sugars tested, 20 g dm-3 glucose was the optimum for shoot regeneration. Rooting of regenerated shoots was successful on 1/4-strength MS medium with the addition of 15.4 M IBA. Almost 100% plantlets survived acclimatization after transferred to soil.Key words: Phellodendron amurense, callus, shoot regeneration, stem explants