Given their remarkable capacity to convert atmospheric nitrogen into plant-accessible ammonia, nitrogen-fixing microbial species hold promise as a sustainable alternative to chemical nitrogen fertilizers, particularly in economically significant crops like wheat. This study aimed to identify strains with optimal attributes for promoting wheat growth sustainably, with a primary emphasis on reducing reliance on chemical nitrogen fertilizers. We isolated free nitrogen-fixing strains from diverse rhizospheric soils across Morocco. Subsequently, we conducted a rigorous screening process to evaluate their plant growth-promoting traits, including nitrogen fixation, phosphate solubilization, phytohormone production and their ability to enhance wheat plant growth under controlled conditions. Two specific strains, Rhodotorula mucilaginosa NF 516 and Arthrobacter sp. NF 528, were selected for in-depth evaluation, with the focus on their ability to reduce the need for chemical nitrogen supply, particularly when used in conjunction with TSP fertilizer and natural rock phosphate. These two sources of phosphate were chosen to assess their agricultural effectiveness on wheat plants. Twenty-two nitrogen-fixing strains (nif-H+) were isolated from various Moroccan rhizospheric soils, representing Bacillus sp., Pseudomonas sp., Arthrobacter sp., Burkholderia sp. and a yeast-like microorganism. These strains were carefully selected based on their potential to promote plant growth. The findings revealed that the application of Rhodotorula mucilaginosa NF 516 and Arthrobacter sp. NF 528 individually or in combination, significantly improved wheat plant growth and enhanced nutrients (N and P) uptake under reduced nitrogen regimes. Notably, their effectiveness was evident in response to both natural rock phosphate and TSP, demonstrating their important role in wheat production under conditions of low nitrogen and complex phosphorus inputs. This research underscores the significant role of nitrogen-fixing microorganisms, particularly Rhodotorula mucilaginosa NF 516 and Arthrobacter sp. NF 528, in wheat production under conditions of low nitrogen and complex phosphorus inputs. It showcases their potential to reduce chemical nitrogen fertilization requirements by up to 50% without compromising wheat plant yields. Our study emphasizes the importance of bacterial biological nitrogen fixation in meeting the remaining nitrogen requirements beyond this reduction. This underscores the vital role of microbial contributions in providing essential nitrogen for optimal plant growth and highlights the significance of biological nitrogen fixation in sustainable agriculture practices.