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Introduction: The Calotropis procera Aiton plant is a valuable and effective medicinal species used to treat various diseases, including as an anti-carminative and for indigestion, as well as for anticancer and anticonvulsant treatments. This species naturally inhabits fragmented habitats in southern Iran, particularly near the coasts of the Persian Gulf and the Oman Sea, in the arid and semi-arid regions of Sistan and Baluchistan, Khuzestan, Bushehr, and Hormozgan. On the other hand, due to the discovery of adverse effects caused by chemical drugs, people have turned their attention towards using medicinal plants. The health of agricultural products, especially medicinal plants, has special importance. The symbiosis of plants with mycorrhiza can be lead to improve of the root development and will affect on the absorption of water and nutrients. Aim of this study was investigate the effects of symbiosis between C. procera Aiton. with three different species of mycorrhizal on some phytochemical and antioxidant activity, and content of potassium, phosphorus, and sodium in shoots of C. procera Aiton. Materials and Methods: This research was conducted in the greenhouse of the Horticultural Science Department, University of Zabol, Zabol, Iran. Transplants were cultured in the soil medium in pot condition. This experiment was carried out in a completely randomized design with four treatments include inoculation with three different mycorrhizal fungi (Glomus intraradicese, Glomus fasiculatum, and Glomus mosseae) and control (without inoculation) and three replicate. Meusered indices were assessed 8 weeks after transplanting. Measured parameters include; growth indices such as fresh and dry weight of root and shoot, and phytochemical indices are chlorophyll a and b, Carotenoid, phenol, flavonoid, antioxidant activity, protein, and concentration of potassium, phosphorus, and sodium in shoots. Data were analyzed by ANOVA test using JMP, and means were compared using the LSD test at P˂0.01. Results and Discussion: The results of variance analysis were shown significant effects of mycorrhizal symbiosis on all investigated indices (P ≤ 0.01). Also, means comparison shows a significant difference between the effects of different mycorrhiza species. The highest content of chlorophyll a and carotenoid was obtained in symbiosis with G. mossae, and the highest content of chlorophyll b existed in symbiosis with G. fasiculatum and G. intraradicese. In this regard, it has been found that mycorrhizal symbiosis leads to an increase in water absorption, and an improvement in the absorption of nutrients by the plant, thereby leading to an increase in chlorophyll synthesis enzymes activity. symbiosis of G. fasiculatum had the highest effects on increasing phenol and antioxidant activity. increase in antioxidant activity leading to strong suppression of reactive oxygen species, which ultimately increases the remedial capability of C. procera Aiton. The highest amount of flavonoid was observed in the condition of symbiosis with G. intraradicese and G. fasiculatum, and the highest content of protein was observed in condition of symbiosis with G. fasiculatum and G. mossae. Symbiosis with the G. intraradicese significantly increased the concentration of potassium and phosphorus in shoots of C. procera Aiton, while symbiosis with G. mossae causes to a significant increase in sodium concentration. Generally, the results of the current study were shown that symbiosis with G. fasiculatum and G. intraradicese are more useful than symbiosis with G. mossae. The symbiosis of plants with mycorrhiza fungi increases the volume and length of their roots, as a result of which the roots have more contact with the soil, and the amount of water and nutrient absorption from the soil increases. Conclusion: results of the current study demonstrate that the phytochemical, antioxidant activity, and concentration of potassium and phosphorus and sodium have differed among extract and shoots related to symbiosis of C. procera Aiton and species of fungi. And symbiosis with G. fasiculatum had the greatest effect in increasing the production of secondary metabolites. [ABSTRACT FROM AUTHOR]