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Lentil is an important annual food legume crop, nitrogen fixer and provides a substantial amount of protein, carbohydrate, minerals, and vitamin content. The use of molecular markers to assess genetic diversity is crucial for crop improvement, efficient management, and conservation of plant genetic resources. The current study aimed to determine the genetic diversity among lentil genotypes using sequence-related amplified polymorphism (SRAP) markers. Therefore, we evaluated a collection of 36 lentil genotypes, including 20 from Yemen, Saudi Arabia (7), Egypt (4), and Bangladesh (3), and (2) genotypes from the International Center for Research in Dry Area (ICARDA) using 21 SRAP primer combinations. The amplified fragments showed a high level of useful polymorphic amplified fragments (775 out of 782) indicating a higher degree of variation. The polymorphic information content (PIC) ranged from 0.31 to 0.39 with an average of 0.33 for each primer. The UPGMA trees, based on Jaccard similarity index matrices, separated the genotypes into four main clusters according to their geographical origin. The population structure supported the major groups and attested to their great degree of differentiation. The highest lentil population was found at K = 3, K = 5, and K = 7 levels, showing purity and admixture ancestry among the lentil population. This study highlighted the wide genetic diversity among the studied lentil genotypes and demonstrated the effectiveness of the SRAP technique in determining the genetic variability of lentil. Furthermore, it could be used to establish the genetic peculiarity of ecotypes when applying for the obtainment of origin and agro-morphological characteristics. [ABSTRACT FROM AUTHOR]