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The effect of Thidiazuron (TDZ), basal media and light quality on adventitious shoot regeneration from in vitro cultured stem of Populus alba×P. berolinensis were determined to establish a high efficiency shoot regeneration system from stem explants of P. alba×P. berolinensis. Stems of Populus alba×P. berolinensis were collected from cultured shoots in vitro derived from dormancy buds of 3-year-old seedlings. The stem explants were cultured on MS medium containing 0.02-mg·L-1 NAA (naphthaleneacetic acid), and 0.1, 0.3, 0.5 and 1.0 mg·L-1 concentrations of TDZ to determine the effect of TDZ on shoot regeneration. Three basal media, i.e. MS, woody plant medium (WPM) and B5, were used to test their influences of different media on adventitious shoot regeneration. Green, red, blue and yellow plastic films in comparison with florescent light as control were used to observe their effects on shoot regeneration. The results showed that different concentrations of TDZ had an evident influence on shoot regeneration. Lower concentration of TDZ (0.1 mg·L-1) resulted in more adventitious shoot regeneration and higher concentration of TDZ (>0.1 mg·L-1) inhibited shoot regeneration. Among different media, MS medium exhibited a high efficiency for shoot regeneration, followed by WPM medium, while B5 medium inhibited shoot regeneration. Normal light and yellow light exhibited better effects on shoot regeneration, compared with other light.