Abstract: In order to clarify the genetic diversity of physical characteristics of flue-cured tobacco germplasm resources and to use germplasm resources effectively, the genetic diversity analysis and cluster analysis were conducted for 6 physical characteristic indexes of C3F leaves of 269 selected flue-cured tobacco germplasm resources. The results showed that: 1) The variable coefficients of 6 physical characteristic indexes of the experimental germplasm resources ranged from 6.73% to 20.25% and genetic diversity indexes were from 1.934 to 2.050 with an average of 2.014. 2) The mean value of genetic diversity indexes for physical characteristics of domestic resources (1.979) was a little higher than that of resources introduced from U.S.A. (1.956) and other resources (1.932), and the mean value of genetic diversity indexes for domestic local germplasm resources (2.628) in China was higher than that of hybrid breeding resources (1.922). 3) The experimental germplasm resources could be clustered into 4 groups at a similar distance of 0.982. There were extremely significant differences in the indexes between the groups. The physical characteristic scores of resources in group Ⅰ and group Ⅱ were higher than those in group Ⅲ and group Ⅳ. The physical characteristic indexes of 7 resources, Fengzi 6, Huangmiaoyu, B22, Gexin 6, Mudan 81-56, black corn cob and Fengcheng gold, were all in the most suitable or suitable range, and their scores were higher. In conclusion, the genetic diversity of physical characteristics of experimental flue-cured tobacco germplasm resources was abundant, that of local resources was higher than that of hybrid breeding resources in China. Seven germplasm resources with excellent physical characteristics could be used for tobacco variety breeding and physical characteristic improving.