Cleome gynandra L. is an important traditional leafy vegetable in Burkina Faso diets. Because of its high nutritional value, it makes a good dietary supplement, providing an important nutritional and medicinal requirement. The aim of the present study was to assess the genetic diversity and intra-and interpopulation relationships of the C. gynandra collection using SSR molecular markers specific to C. gynandra. Nine microsatellite primers were used to assess the genetic diversity of 33 C. gynandra accessions. A total of 50 alleles were counted (an average of 5.56 alleles per marker). Polymorphism information content (PIC) ranged from 0.82 to 0.43 for markers CG01 and CG033, respectively, with an average of 0.60, showing the high genetic diversity of the C. gynandra collection studied. In fact, six markers are highly informative (PIC > 0.5). Inter-morphotype differentiation showed an STF of 0.17 was recorded between the green morphotype and the dark purple morphotype. In effect, the two morphotypes are different. The organization and structuring of diversity resulted in three distinct genetic groups, irrespective of morphotype color, and accession origin.