A study was carried out to compare the effects of treating wheat (Triticum aestivum) and Italian ryegrass (Lolium multiflorum) with atrazine and fluorodifen. The herbicides interfered with photosynthesis and dark respiration, depending on the species. Atrazine decreased photosynthesis in both species and dark respiration in wheat, while fluorodifen caused decrements of photosynthetic activity of wheat. Antioxidant enzymes, such as ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), and glutathione reductase (GR), were generally more active in untreated and treated wheat with respect to Italian ryegrass, which explains why oxidative damage, expressed as malondialdehyde (MDA) content, was only found in ryegrass. Investigations on the activity of herbicide-detoxifying enzyme, glutathione S-transferase (GST), and on the accumulation and persistence of the herbicides in the plants showed higher detoxification rates in wheat than in the grass.