Abstract:This paper describes a novel method based on etiolation treatment for micropropagation of Populus tomentosa. It includes: (1) Antibiotic “cefotaxime” (2.5 ppm in nutrient medium) is used to inhibit bacteria growth and the cultures which have been contaminated after long period subculture are recovered by sterilization with 1/10000 HgCl2(w/v). (2)Low concentration of TDZ (thidiazuron, 0.005 ppm) replaces expensive zeatin for enhancing bud differentiation and multiplication of leaf explants. (3) Shoot elongation is promoted after etiolation treatment, which will increase the number of shoots suitable for rooting, about 50 etiolated shoots are obtained in a 100 ml flask, 3—5 times more than those produced in traditional method. (4) Etiolated shoots or after their greening are used as cuttings for rooting in vivo and over 90% survival rate could be achieved when the medium is sterilized. This method is labour and money saving and high efficient.