Abstract:With semi-lignified shoots sprouted from the girdling of 13 years old Quercus shumardii as explants, we established the regeneration system of Q.shumardii by in vitro culture. When the WPM was supplemented with 1.0 mg·L-1 BA , 0.1 mg·L-1 NAA and 5 mg·L-1 Biotin, 5 mg·L-1 acetylcholine, 5 mg·L-1 vitamine B2, and cultured for 40 d, the multiplication coefficient was 2.15. The rooting rate of single bud was 10% when cultured in 1/2 WPM supplemented with 2.0 mg·L-1 NAA, 5 mg·L-1 Biotin, 5 mg·L-1 acetylcholine, 5 mg·L-1 vitamine B2. The rooting rate was about 80% when roots were induced out-tube. Our work established technical system for tissue culture and plantlet regeneration of Q.shumardii with a solid foundation for the extension of its good quality seedlings.