The composition and quantity of Restorer-of-fertility genes contained in the restorer line directly determine the strength of the restoration ability, thereby affecting the fertility stability of hybrid soybeans bred through the "three line" method. The Rf1 gene is widely present in soybean cytoplasmic male sterile restorer lines, while Rf2, as a newly discovered strong Restorer-of-fertility gene, only exists in a very small number of restorer lines. To this end, this study developed functional molecular marker for the Rf2 gene and used conventional hybrid breeding combined with molecular marker-assisted breeding techniques to aggregate the Restorer-of-fertility genes Rf1 and Rf2, creating a new germplasm of soybean cytoplasmic male sterile restorer lines with strong restoration ability and excellent agronomic traits. The results indicate that the combination of Rf1 and Rf2 molecular markers can quickly and accurately screen target Restorer-of-fertility gene types, significantly improving the breeding efficiency of new germplasm of gene aggregation restorer lines. At the same time, the newly created restorer germplasm showed excellent performance in agronomic traits such as pod number per plant, grain number, and 100-seed weight. This study provides excellent new germplasm for the breeding of "three line" hybrid soybeans, and also provides technical support for the efficient breeding of more strong restorer lines in the future