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Iron toxicity stress causes physiological and metabolic changes in rice and other plants. To date, there is little information about the metabolite profile of rice under Fe toxicity conditions. In fact, metabolite has a contribution to the physiological condition of plants. Plant metabolomics is a study of low-molecular weight metabolites in plants under certain conditions. The objective of the research was to investigate physiological and metabolic changes in rice under Fe toxicity stress. Two-week-old seedlings of four rice varieties with various Fe toxicity tolerance levels were stressed hydroponically with 400 ppm FeSO 4 . 7H 2 O for 10 d. Numerous physiological characters were observed and untargeted metabolomic analysis was carried out using gas chromatography-mass spectrophotometry (GC-MS). The results showed Fe toxicity induced physiological and metabolite variation in rice. By comparing the metabolites synthesized in Fe toxicity-stressed plants with control plants, it showed that elaidic acid, linoleic acid, and linolenic acid could be as metabolite marker candidates for rice response to Fe toxicity stress. When plants exposed to Fe toxicity stress, elaidic acid increased, whereas linoleic- and linolenic acid decreased. The alteration of fatty acid composition in the root and shoot suggests the alteration of metabolites is one of the tolerance strategies of rice to Fe toxicity stress. This finding offers an insight about the tolerance strategies of rice under Fe toxicity stress related to the maintenance process of the cell membranes during this stress. The genes underlying biosynthesis of the fatty acid could be a target of future research on responsible genes for Fe toxicity tolerance in rice.