Phoenix dactylifera L. is a flowering plant species commonly known as the date palm and is widely cultivated in most Middle East countries, including Libya. The present study analysed the biochemical and genetic diversity of fully mature eight Libyan date palm cultivars grown in different regions using the amplified fragments length polymorphism (AFLP) technique. Six pairs of AFLP molecular marker combinations were utilised to discriminate the eight date palm genotypes. Fruit dimensions (length x diameter) varied based on the type; Majhool Alheelo fruit had the highest value (15.29 cm2), while the lowest value was for Alkhadraya fruits (7.9 cm2). Reducing sugar content ranged from 10.4 per cent of flesh dry weight in Umfetity cultivars to 61.2 per cent in Sufeer-genab, which also showed the highest polymorphism percentage (P%=4.9), while Alkhadarya was the lowest (P%=0.519). The phylogenetic tree indicated that the most distantly related cultivars were Sufeer-genab, Alhamraya and Majhool Alhelo. The two most closely related cultivars were the Alsaeedy Show and Alkhadarya, grown in different regions. Our results indicate that the nutritional and genetic diversity of Libyan cultivars is not closely matched with the growing region