This study was conducted at the Department of Genetics, Faculty of Agriculture, Sohag University to isolate and identify 
the non-rhizobial nodule-associated bacteria at molecular level from different leguminous crops (Egyptian clover, Peanut and Lupine) in Sohag Governorate. A total of 25 nodule-associated bacteria were isolated during this study. All 25 isolates were characterized using symbiotic (nodA) gene loci which found normal in Rhizobium sp. It was found that all isolated bacteria roduced clear PCR fragments of approximately 666 base pairs (bp) except the seven isolates (1, 9, 10, and 17) of Egyptian clover and (19, 21 and 22) of Lupine. Among the seven non-rhizobial nodule-associated bacterial isolates, three isolates (no., 1 and 9 from Egyptian clover and no. 21 from Lupine) were identified by 16S rRNA gene sequencing analysis. These three isolates were symbolized as EMC, EME and EM7, respectively. The obtained data indicated that all of the bacterial isolates were belonging to the genus Pseudomonas. It was found that the two isolates, EME and EM7 gave 100% of similarity with Pseudomonas lini strain DLE411J, NR_029042, and Pseudomonas viciae strain 11K1, NR_180948, respectively. While, the third isolate (EMC) gave 97.72% of similarity with Pseudomonas lini strain DLE411J, NR_029042. The phylogenetic tree based on the 16S rRNA gene sequence of the three isolates supported the obtained results. 
Pseudomonas genus is commonly found coexisting with rhizobia in leguminous crop nodules. It could be used as a single bioinoculant or in combination with rhizobia to improve nodulation and plant growth of leguminous crops.