<p>This study analyzed the fertility and genetic diversity of 75 polyembryonic sugar beet samples that had been improved with the introduction of foreign varieties, aiming to screen outstanding polyembryonic germplasm at the molecular level and provide support for subsequent breeding efforts. The main findings of the study are as follows: Among the 75 single-plant DNA samples, cytoplasmic fertility detection using the TR1 primer indicated that 62 accessions possessed the N-type (normal fertile cytoplasm), accounting for 82.67%. When cytoplasmic fertility-related genes were detected using the S17 primer, no amplification was observed in two samples, while 69 accessions showed amplified bands of 1800&#xa0;bp, representing 92% of the total. Double digestion analysis revealed that the frequency of germplasm resources with non-restored Rf1 allelotypes was very low. In the polyembryonic pollinator line resources of sugar beet, the proportion of maintainer lines is extremely low. To assess genetic diversity, 12 CEAP (Cis-element amplified polymorphism) primers and 8 SRAP (Sequence-related amplified polymorphism) primers were employed, generating a total of 81 polymorphic bands. The UPGMA cluster analysis results showed an average genetic distance of 0.287, and the 75 polyembryonic germplasm resources could be divided into six distinct groups. The results of this study provide a solid foundation for expanding the scope of germplasm resource screening and selecting sugar beet lines with excellent traits through subsequent field experiments.</p>