【Objective】Osmanthus fragrans is one of the ten traditional famous flowers in China. However, due to its short flowering period and small differences in flower type and color, it is difficult to identify different Osmanthus varieties through the traditional morphological strategy. In this study, SSR primers were developed based on the whole genome sequence of O. fragrans, which provides markers for variety identification and genetic diversity analysis of O. fragrans.【Method】Utilizing the published whole-genome data of O. fragrans, genome-wide SSR loci were systematically identified using MISA software. The distribution patterns, density, and motif characteristics (repeat unit length, frequency, and type) of SSR loci were comprehensively analyzed. Primer 3 software was then employed to design primers targeting polymorphic SSR regions. Subsequently, the polymorphism and stability of 131 pairs of primers were further verified. 【Result】A total of 180 838 SSR loci were obtained from the whole genome sequence of O. fragrans, with an average of one locus per 4.10 kb. Mononucleotide (42.59%), dinucleotide (39.03%), and trinucleotide (14.13%) repeats dominated the SSR types. Dinucleotide motifs were primarily AT (49.72%) and AG (46.68%), while trinucleotide repeats were predominantly composed of AAT (28.67%), ACC (18.86%), and AAG (13.37%). The 131 pairs of primers were sequenced by capillary electrophoresis in 32 varieties of four groups of Albus Group, Luteus Group, Aurantiacus Group and Asiaticus Group, and 16 pairs of primers were polymorphic; the number of alleles at a single locus was 4-16, and the average number of alleles at a single locus was 8.94.【Conclusion】The 16 newly developed SSR markers exhibit high polymorphism and reliability, providing an effective molecular toolkit for cultivar authentication, genetic diversity assessment, and fingerprint database establishment in O. fragrans.