The measurements and tests of the samples conducted in the laboratories of the College of Agriculture included isolating bio-fertilizers and testing the efficiency of isolates that fix atmospheric nitrogen and solubilize phosphorous compounds. Bacteria were isolated and identified from the rhizosphere soils of different plants collected from various agricultural areas. A total of 74 bacterial isolates were obtained based on the phenotypic characteristics of the developing colonies, as well as biochemical and microscopic traits. The results of isolation and identification showed that among the 74 bacterial isolates, there were 15 isolates of A. chroococcum, 13 of Az. lipoferum, 13 of B. megaterium, 10 of P. putida, 10 of Actinomycetes, and nitrifying bacteria (Nitrosomonas and Nitrobacter) with 7 and 6 isolates respectively. Bacillus isolates demonstrated the ability to dissolve phosphate compounds with a dissolution diameter ranging between 6-11 mm. The results of the polymerase chain reaction (qPCR) examination confirmed the diagnostic results using the quantitative polymerase interaction technique after extracting DNA from bacterial isolates from the soil. The first group of 10 isolates of P. putida showed, when the gene (GltA F, GltAR) was amplified, resulting bundles with a molecular size of 288 base pairs for P. putida bacteria, confirming the diagnosis using molecular, biological, microscopic, and phenotypic tests. This match confirms the accuracy of the classification of P. putida bacteria. The second group of 10 isolates of P. putida showed, when the gene (PR o DF, PRp o DR) was amplified, resulting bundles with a molecular size of 478 base pairs for B. megaterium, confirming its classification. The third group of 10 isolates of bacteria showed, upon gene amplification (N i f HF, N i f HR), resulting bundles with a molecular size of 371 base pairs for A. chroococcum bacteria, confirming their identification. The fourth group of 14 isolates showed, when the gene (16sAZ F and 16sAZ R) was amplified, resulting bundles with a molecular size of 646 base pairs for Az. lipoferum bacteria, confirming its classification. The fifth group did not show any type of nitrogen-fixing bacteria or phosphate solubilizers after amplification, based on the Sr DNA 71 genotype sequence