A total of 105 transgenic rice lines independently transformed with a pea ferritin gene (Fer) were previously obtained. After seven generations of selfing and β-glucuronidase (GUS) assisted selection, 82 transgenic lines with stable agronomic traits were got. Among the 82 transgenic lines, two high-iron transgenic rice lines Fer34 and Fer65, with the iron contents in the milled rice being 4.82 and 3.46 times of that of the wild type Xiushui 11, respectively were identified. In the two transgenic lines, the exogenous Fer gene was highly expressed, and inherited as a single locus. The transgene had no negative effect on the agronomic traits of rice plant, other mineral nutritional components, appearance quality and eating quality of the milled rice, indicating that these two lines were elite high-iron breeding lines. Furthermore, the practical application and further studies facilitating utilization of the two elite breeding lines were discussed. After the seven generations of selfing and β-glucuronidase (GUS) assisted selection, 82 transgenic lines with stable agronomic traits were got. Among the 82 transgenic lines, two high-iron transgenic rice lines Fer34 and Fer65, with the iron contents in the milled rice being 4.82 and 3.46 times of that of the wild type Xiushui 11, respectively, were identified. In the two transgenic lines, the exogenous Fer gene was highly expressed, and inherited as a single locus. The transgene had no negative effect on the agronomic traits of rice plant, other mineral nutritional components, appearance quality and eating quality of the milled rice, indicating that these two lines were elite high-iron breeding lines. Furthermore, the practical application and further studies facilitating utilization of the two elite breeding lines were discussed.