The Apiaceae family includes a few agronomic and medicinal species, one of which is Scaligeria lazica Boiss. In this study, the genetic diversity of S. lazica was analyzed based on novel simple sequence repeat (SSR) markers using next-generation sequencing (NGS). A total of 15.17G clean Illumina data set was obtained and dinucleotide repeats were the most abundant repeats in S. lazica. Of the tested 150 SSR primer pairs, 139 ones produced amplification and 84 ones were polymorphic. Forty polymorphic SSR loci were used in genetic diversity analysis of 40 S. lazica accessions from four locations. A total of 264 alleles were amplified with an average of 6.6 alleles per locus. The polymorphism information content (PIC) was 0.60, while the observed homozygosity (Ho) and expected heterozygosity (He) values were 0.47 and 0.66, respectively. According to cluster and structure analysis, all accessions were grouped into four different clusters according to their collection sites. The SSR markers developed in this study can be tested for other Scaligeria species due to their high transferability and can be used for genetic studies in genus Scaligeria DC.