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An effective in vitro regeneration system is one of the prerequisites for genetic manipulation of alfalfa (Medicago sativa L.) varieties and genotypes. In this research, somatic embryogenesis of four alfalfa genotypes, 6-18 (synthetic), 4-14 (Kara Yonje- Karakozlu), 3-27 (Kara Yonje Maraghe) and y-6 (Regen-SY), were investigated using leaf and petiole explants. Formation of callus and somatic embryogenesis was significantly influenced by the explant type and interaction of genotype and culture medium. Petiole explants of genotype 4-14 produced the highest yield of callus (0.406 gr fresh weight of callus). Percentage of somatic embryogenesis and the number of embryos per callus in petiole explants of genotype 4-14 was higher than those of other genotypes and explants. In genotype 6-18, the highest percentage of somatic embryogenesis was achieved on MS medium containing 5 mg/L 2,4-D and 2 mg/L kinetin. There was no significant differences between genotypes and explants in terms of embryo conversion to plantlet, and on average, 58% of somatic embryos converted to plantlet on MS medium. The petiole explants of genotype 6-18 did not exhibit somatic embryogenesis response in medium containing low ratio of 2,4-D:Kinetin (5 mg/L 2,4-D and 2 mg/L kinetin). While, these explants showed somatic embryogenesis in higher ratio of 2,4-D:Kinetin (5:1). The plantlet conversion efficiency of somatic embryos produced through this study was relatively higher and therefore, the method presented in this study could be used in alfalfa genetic manipulation and molecular studies. [ABSTRACT FROM AUTHOR]