Since the global approach to the production of medicinal plants is effective in improving the quantity and quality of the material, it seems that the nutrition of these plants through the application of biological fertilizers in different environmental conditions is most in line with the production goals of medicinal plants. Therefore, in order to investigate the effect of bio and non-bio fertilizers on biochemical and morphological characteristics of Peppermint (Mentha piperita) under different irrigation intervals, an experiment in the form of a split-plot in a randomized complete design with 4 replications in the research farm of Agricultural and Natural Resources Research Center, Yazd, Iran was conducted in 2020. Experimental factors include three irrigation intervals: Control (every 7 days), every 12 days, every 17 days, and different levels of fertilizer including Control (without fertilizer application), mycorrhizal (Glomus), Azperium brasilense, Pseudomonas fluorescens, and chemical fertilizer (NPK). The results showed that with increasing irrigation intervals, height decreased by 33%. Comparison of the mean of interaction effects of the treatments showed that the biological yield decreased by 81% compared to the control treatment. Also, the highest rate of biological yield was observed in the the7-day irrigation interval and mycorrhiza fertilizer. Comparison of the mean of the data showed that with increasing the irrigation interval, the percentage of essential oil increased. Also, the use of mycorrhiza had the highest percentage of essential oil (2.54%). Therefore, the use of bio-fertilizers, especially mycorrhiza which can greatly reduce the adverse effects of peppermint in dehydration stress conditions is recommended.