Genetic diversity among 70 maintainers and two pollinators of sub-Saharan and Indian origin was studied for simple sequence repeat (SSR) loci using 34 primer pairs. A total of 213 alleles were detected with an average of 6.26 alleles per locus. Polymorphic information content (PIC) ranged from 0.05 to 0.96 with a mean of 0.58 for the SSR loci. Mean PIC across the linkage groups and number of alleles in dinucleotide motifs varied significantly. The lowest PIC (0.239) for linkage group 6 indicated comparatively conserved nature of this linkage group. Genetic similarity estimates ranged from 0.05 to 0.73 with an average value of 0.29. This indicated sufficient diversity among the maintainer and pollinator lines. The 72 lines fell in five clusters, and the clustering pattern corroborated with their pedigree and characteristic traits. Pollinator ICMR 356 was more diverse from the maintainer lines analysed, and can be a potential parent for pearl millet hybrid development.