Backcrossing is a trait introgression method of renewed importance in crops. The evolution of introgressed or substituted segments through backcross generations has been analyzed theoretically using simulations. In this study, the content of donor genomes, including donor segment number (DSN), donor segment length (DSL), and donor genome size (DGS), were directly analyzed in six crosses over three successive backcrosses using a set of single sequence repeat (SSR) markers covering the entire rice genome. The results of this analysis demonstrated that the average DSN in each genome was 8.39 in BC₂F₁, 4.13 in BC₃F₁, and 2.41 in BC₄F₁, decreasing nearly by half with each backcrossing. The average DSL was 33.43 cM (centiMorgans) in BC₂F₁, 29.04 cM in BC₃F₁, and 25.07 cM in BC₄F₁, display a progressive decrease slightly greater than 10% in each additional backcross generation. Meanwhile the average DGS was 280.51 cM in BC₂F₁, 119.97 cM in BC₃F₁, and 60.53 cM in BC₄F₁, decreasing 57.2% from BC₂F₁ to BC₃F₁ and 50.4% from BC₃F₁ to BC₄F₁. This revealed that the reduction in DGS was approximately 50% with each backcrossing. These results provide a guide for introgression or substitution of target chromosome segments from donors into recipients in backcross programs.