Five Tunisian varieties of Kabuli chickpea were characterized based on agro morphological, molecular and biochemical parameters to investigate their genetic variability and yield potential. Randomized complete block design field trials were carried out in the upper semi-arid region of Kef in Tunisia during the 2013-2014 seasons. Data analysis showed significant differences between genotypes for several parameters. The results indicated that these genotypes could be set into two different groups. The first group composed of Bochra and Chetoui genotypes. Kasseb, Neyer and Beja1 were in the second group. Genotypes in each group were closely related to each other according to their common morphological characters such as pod number, one hundred seeds weight and yield. Chetoui and Kasseb Journal of Agricultural StudiesISSN 2166-0379 2017, Vol. 5, No. 1 http://jas.macrothink.org 78 varieties are later in comparison to other varieties. Genetic diversity was studied using simple sequence repeat (SSR) markers. Four loci (TA64, TA71, TA96, TA194) were multiallelic. Whereas while two loci (TA72, GAA47) were monomorphic. Polymorphism analysis showed a phylogeny related to genotypes differentiation according to their relatives, origin and several morphological characters. Bochra variety had high amino acids content followed by Chetoui variety. All the varieties were deficient in sulfur amino acids. Chickpeas protein contents were variable and high ranging from 18% to 25%.