<b>Background and Objective:</b> Rhizobia are bacteria including genes codes for enzymes involved in the fixing of the atmospheric nitrogen. A set of twenty rhizobial isolates were studied to determine their morphological, biochemical, molecular characteristics using the 16S rRNA gene in addition to assess their growth and symbiotic performance. <b>Materials and Methods:</b> Rhizobial isolates were isolated from root nodules of <i>Vicia faba </i>L. plants. The isolates were morphologically characterized by determining cell shapes, size, Gram stain reaction, motility, sporulation, bacterial growth performance was determined by IAA production and biomass density. Symbiotic performance was measured by evaluation of nodulation status and shoot/root dry weight. Sequencing of 16S rRNA and phylogenetic analysis were done for the five promising isolates. Statistical analysis was performed using a one-sample Student t-test. <b>Results:</b> Only five rhizobial isolates (Rh 32, Rh 6-A, Rh 3-4, Rh RL3 and Rh 8-A) were selected according to their growth and symbiotic performance and subjected to further molecular characterizations. All isolates were found to have remarkable nodulation status, IAA production, nitrogenase activity and increasing the root and shoot dry weight. The five selected rhizobial isolates were identified by partial sequencing of 16S rRNA genes and registered in the GenBank database. The alignment and phylogenetic analyses of 16S rRNA sequences closely related in the GenBank revealed that all isolates belonging to <i>Rhizobium leguminosarum</i> bv. viciae. <b>Conclusion:</b> The results confirmed that the five Rhizobial strains will be promising as a source of genes for nitrogen fixation and plant growth promotion.
Keywords: 16S rRNA gene; Rhizobium; chemical fertilizers; faba bean; genetic diversity; nitrogen fixation; phylogenetic; sequencing.