Water deficit is one of the most important environmental stresses that adversely affect crop growth and production and mycorrhizal fungi and symbiotic bacteria have important role in resistance to drought stress. The effect of biofertilizers on alfalfa stress tolerance was studied at the greenhouse condition. Treatments comprised three water-deficit stresses (35%, 55% and 75% of field capacity) and four seeds inoculations (Glomus mosseae, Sinorhizobium meliloti, G. mosseae + S. meliloti and non-inoculated). Water-deficit stress decrease cell membrane stability (39%), total Chl (24.05%), carotenoid (35.55%), quantum yield (50.64%) and forage yield (28.20%), while increased the proline and soluble sugars content (68.55 and 46.53% respectively) and osmotic potential (45.84%). The inoculation of seeds increased the capability of the plants in counteracting the stress, so that the production of compatible solutes was increased and the photosynthetic indices, proline, osmotic potential, membrane stability and forage yield were improved by seed inoculation. Mycorrhiza improved photosynthetic indexes and proline, but bacteria had more efficacy on membrane stability and forage yield. However, double inoculation due to the synergistic effect of mycorrhiza and Sinorhizobium, had the greatest effect than Solitary inoculation. Our results suggest that biofertilized alfalfa plants were better adapted than non-biofertilized ones to cope with water deficit.