Little information is available on the role of glycinebetaine (GB) in chromium (Cr) tolerance while Cr toxicity is widespread problem in crops grown on Cr-contaminated soils. In this study, we investigated the influence of GB on Cr tolerance in wheat (Triticum aestivum L.) grown in sand and soil mediums. Three concentrations of chromium (0, 0.25, and 0.5 mM) were tested with and without foliar application of GB (0.1 M). Chromium alone led to a significant growth inhibition and content of chlorophyll a, b, proteins and enhanced the activity of antioxidant enzymes. Glycinebetaine foliar application successfully alleviated the toxic effects of Cr on wheat plants and enhanced growth characteristics, biomass, proteins, and chlorophyll contents. Glycinebetaine also reduced Cr accumulation in wheat plants especially in grains and enhanced the activity of antioxidant enzymes in both shoots and roots. This study provides evidence that GB application contributes to decreased Cr concentrations in wheat plants and its importance in the detoxification of heavy metals.