In this experiment, the effect of inoculation of wheat cultivars with Azospirillum brasilense and pH of irrigation water on yield, protein content and sedimentation rate of protein (SRP) for wheat grain were examined. For this reason, a factorial experiment was conducted with three replicates and three cultivars of wheat (Omid, Ghods and Roshan), which were inoculated with and without Azospirillum brasilense. Irrigation water was applied into the pots during growing season with different levels of pH (7, 9 and 11). At the harvest time, the yield (grain) and nitrogen, protein content and SRP in grain were measured. The results indicated that the seed inoculation, pH of irrigation water and wheat cultivars have significant effect on the yield, protein content and SRP. Increase in the pH of irrigation water caused to decrease the yield, protein content and SRP. The maximum decline in the yield (17.2% decreases due a unit increase in pH), protein content and SRP due to the increase in irrigation water pH was observed in the Omid wheat cultivar (most sensitive cultivar), in contrast the Roshan wheat cultivar shows less adverse effect (10/7% decreases due a unit increase in pH). In inoculated treatments, all measurements were higher than non-inoculated pots. According to the results, Azospirillum can modify unfavorable conditions (higher pH) to obtain higher yield (by 10/7%), protein content (by 15.5%) and SRP (by 7.9%). However, the adverse effect of pH of irrigation water was the same in inoculated plants. From statistical point of view, the interaction of Azospirillum, wheat cultivar and pH of irrigation water had a significant effect on yield (grain) however the interaction for protein content and SRP were not significant. The interactions of each two treatments had statistically significant effect on yield, protein content and SRP; thereby more attention is needed for selection of Azospirillum stain and wheat cultivar.