Amplified fragment length polymorphism (AFLP) markers and flow cytometry were applied for the genetic characterization of wild and cultivated accessions of Sesamum radiatum and Cerathoteca sesamoides; two neglected and underutilized species of traditional leafy vegetable consumed in Benin. The average 2C nuclear DNA content per nucleus was found to be 1.99 ± 0.06 and 1.05 ± 0.06 pg for S. radiatum and C. sesamoides, respectively which correspond to estimated genome size of 1946,22 Mpb for S. radiatum and 1026,9 Mpb for C. sesamoides. No variation in DNA content could be detected within accessions from each analysed species. Also, no relation was found between nuclear DNA content, ecogeographical origin and the status (cultivated or wild) of the analyzed accessions. AFLP markers revealed low diversity within the accessions analyzed. Results from the study contributed to a better characterization of S. radiatum and C. sesamoides accessions and will help in defining both genetic resources conservation and breeding strategies.