Abstract:Several factors affecting transformation of Populus tomentosa Carr. were studied, and a simple and effective protocol with optimized condition for transformation of P. tomentosa was developed. The results demonstrated that the transformation frequency was extremely increased with the presence of acetosyringone, and likely with the Agrobacterium tumefaciens (Smith et Townsend) Conn density, duration of infection and co-cultivation. It was found that removal of CoCl2·6H20 from the co-culture medium was benefitial to obtain Kanr shoots.