Phytohormones and fatty acids play a significant role in developmental stages of plant growth and defense against biotic and abiotic stresses. The purpose of this study was to determine the spermidine (Spd)-induced phytohormones and fatty acids changes involve the acclimation of cucumber plants against salt stress. Plants exposed to salt stress had significant reduction in their growth. Exogenously applied Spd increased the shoot length and protein content in salt-stressed plants. The accumulation of total phenol and malondialdehyde was higher in salt-affected plants than in their controls and these detrimental effects were mitigated by Spd treatment. Moreover, salt stress caused a significant increase in salicylic acid (SA) and jasmonic acid (JA); while Spd treatment ameliorated these salt stress effects by reducing SA and JA content. The marked accumulation of total free fatty acid was observed in salt-stressed plants, while the application of Spd to salt-stressed plants reduced the total free fatty acid content. In addition, Spd inhibited the stearic acid, linoleic acid and linolenic acid in salt-stressed plants. The results of current study suggest that exogenous application of Spd-induced phytohormones and fatty acids changes would be a reason for increasing the acclimation of cucumber plants under salt stress condition.