To evidence a possible mechanism of defense toward oxidative stress induced by herbicides in plants, an investigation was carried on the activity of glutathione peroxidase (GPX) in Italian ryegrass (Lolium multiflorum) and in Festuca (Festuca arundinacea) in response to atrazine (6-chloro-N-ethyl-N'-isopropyl-1,3,5-triazine-2,4-diamine) and fluorodifen (4-nitrophenyl α,α,α-trifluoro-2-nitro-p-tolyl ether). In general, the herbicide treatments significantly induced GPX activity in the shoots of Italian ryegrass, whereas inhibited it in Festuca. These opposite behaviors are examined, taking into account the accumulation and persistence of the two herbicides in the plants, and they are discussed in terms of GPX counteraction to oxidative stress in the first case, and of a lower detoxification rate unable to prevent a deleterious effect on the GPX activity in the second case. Further information on the properties of Italian ryegrass and Festuca GPX were achieved by purification and isolation of the enzymes, performed by protein liquid chromatography and by electrophoretic analyses. GPX of both the plants were found to be heterodimer with multiple function in showing also glutathione S-transferase (GST) activity.