Nowadays, the use of soil-born microorganisms as biological fertilizers is considered to be a natural and most desirable solution to maintain sustainability of agricultural soil system. Potassium releasing bacteria, nitrogen fixing and phosphorus dissolving bacteria make mentioned elements available to plants. In order to evaluate the effects of bio-fertilizers Potabarvar 2, Sinorhizobium meliloti, as well as urea fertilizer on physiological properties and yield of Fenugreek, an experiment as complete randomized block design was conducted with five treatments and three replications. Treatments included biofertilizer Potabarvar 2, S. meliloti, inoculation with a mixture of Sinorhizobium+Potabarvar 2, positive control (based on soil analysis) and negative control (no fertilization and inoculation).The results showed that all morphological traits were significant at 1%. Most physiological traits except for carotenoid were significantly affected by S. meliloti, and a mixture of Sinorhizobium+Potabarvar 2. Seed inoculation with biofertilizer Sinorhizobium meliloti and Potabarvar 2 lead to increase in growth and eventually shoot yield. Separate application of these biofertilizers led to better results than the integrated application. Symbiotic relationship of Sinorhizobium with Fenugreek increased physiological indices data, especially the absorption of nitrogen and phosphorus, as well as the amount of phenolic antioxidant have been significantly affected. In general, application of S. meliloti resulted in better and more effective increase in yield, quality and plant growth than fertilizer Potabarvar 2 and a mixture of Sinorhizobium+Potabarvar 2.