Sesame (Sesamum indicum L.) is an important crop in tropical and subtropical areas. Sesame seed is traditionally used for direct consumption, because of its high nutritional value (50 % oil and 25 % protein), and as a source of oil of excellent quality. Potentially beneficial effects of sesame on human health have recently renewed the interest in this ancient crop.Information on the genetic diversity in sesame is limited, only some studies about morphological diversity have been carried out, and generally, these studies have been focused on regional interest. To overcome this gap of knowledge in sesame, this research was achieved to know how diverse is a sesame germplasm collection containing both accessions from different origin regions, and commercial cultivars or experimental lines. Genetic diversity was assessed at three different levels: DNA (by means of amplified fragment length polymorphism or AFLP), metabolic profiles, and functional metabolites (by means of effect of plant extracts on soil-borne pathogenic fungi). Individual outputs for every approach were obtained, but also information about usefulness of AFLP for identifying sesame cultivars, optimal conditions for bioassays, and identification of sesame accessions potentially valuables for breeding and/or production of natural compounds against soil-borne fungi The assessment by means of AFLP resulted in a high level of variability within all diversity centres except Central Asia. No association between geographic origin and AFLP patterns was found. Most