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The objective of this paper was to investigate the protective effect of three potential herbicide safeners (3-dichloroacetyl oxazolidine and its two optical isomers) on detoxifying to chloroacetanilide herbicide acetochlor in maize. In this study, physiological and biochemical tests were conducted under laboratory condition in 2015. All safeners increased the expression levels of herbicide detoxifying enzymes, including glutathione S-transferases (GST), catalase (CAT) and peroxidase (POD) to reduce chloroacetanilide herbicide phytotoxicity in maize seedlings. Our results suggest that the R-isomer of R-29148 can induce glutathione (GSH) expression, GST activity, and affinity for the 1-chloro2,4-dinitrobenzene (CDNB) substrate in maize, which can protect maize from injury by chloroacetanilide herbicide acetochlor. Further information on the chiral safener role in antioxida