Aerobic organisms need antioxidant defense systems to deal with free radicals which either are produced during aerobic respiration or may have an external origin. Oxidative stress, which is resulted from an imbalance between the production of free radicals and the ability of antioxidant defense mechanism to deactivate them, is involved in the development of many chronic diseases such as cancer, diabetes, CVD and some neurodegenerative diseases. Reinforcing the antioxidant potential of the body has been considered as a strategy that could prevent and manage such conditions. In the previous review article published by British Journal of Nutrition, in 2014, for the first time, we concluded that riboflavin could alleviate oxidative stress. Although riboflavin can serve as a prooxidant when exposed to ultraviolet irradiation, the literature is replete with studies that support its antioxidant properties. Furthermore, recent evidence suggests that riboflavin may have a therapeutic potential in many conditions in which oxidative stress is involved, although the therapeutic efficacy of riboflavin as an antioxidant requires further study under conditions of wellness and clinical disease.

Out of 1,343 mutant lines of rice mutated by sodium azide from the parental Japonica-type variety ‘Tainung 67’ (TNG67), a paraquat-susceptible line 1192 and a paraquat-tolerant line 72 were selected using whole seedlings at the four-leaf stage and leaf segments at the tillering stage as test materials. Further selection from progenies of these two mutant lines yielded the susceptible 1192-11 (S) and tolerant 72-16 (T), which were studied herein. Chlorophyll fluorescence, electrolyte leakage, and lipid peroxidation were measured for leaf segments of rice following treatment with 0.1 mM paraquat. A comparison of these responses among the three rice lines (TNG67, 72-16, and 1192-11), revealed a higher tolerance to paraquat in the tolerant mutant line 72-16 and the parental variety TNG67 than in the susceptible mutant 1192-11. Analysis of the antioxidative system in paraquat-treated leaf segments showed that the reduced form of glutathione (GSH) and the ratio of GSH to total glutathione increased by 3.5-fold within 6 h after treatment (HAT) and up to 5-fold 9 HAT in the T line, as compared with the S line. In view of the high activities of both dehydroascorbate reductase (DHAR) and glutathione reductase (GR) in paraquat-treated leaves of TNG67 and the T line, the antioxidative effect of the ascorbate–glutathione cycle is hereby proposed to play an essential role in paraquat tolerance of rice. Pretreatment of rice segments with spermine enhanced DHAR and GR activities as well as paraquat tolerance of the S line. These results suggest that the activity of ascorbate–glutathione cycle induced by spermine is involved in rice tolerance to this herbicide. Although kinetics studies showed no significant difference among the three rice lines in paraquat inhibition of GR, a lower affinity of enzyme to substrate (Km) in TNG67 and the T line and a higher maximal reaction rate (Vmax) in the T line for the oxidized glutathione substrate (GSSG) were detected. These observations further implicate the importance of glutathione reductase in paraquat tolerance of rice.

In order to explore the physiological mechanism of paraquat resistance in tall fleabane, a widespread weed in Taiwan where resistance to this herbicide has been observed since 1980, the role of the antioxidative system was assessed. The susceptible (S) and resistant (R) biotypes of tall fleabane were distinguished clearly by the relative distribution frequency of injury index caused by 78 µM paraquat. Although malondialdehyde, an indicator for peroxidation damage to the plant, in the the R-biotype was not changed, in the S-biotype malondialdehyde increased within 2 h after treatment of 50 µM paraquat. Analysis of several antioxidants and pertinent enzymes revealed that ascorbate peroxidase activity was decreased by paraquat treatment in the S-biotype; and a lower basal level of ascorbate was present in the S-biotype as well. The maintenance of a high ratio of reduced glutathione to total glutathione, coupled with a pronounced and rapid increase of glutathione reductase (GR) activity in the the R-biotype, suggests that an active reduced glutathione/oxidized glutathione (GSH/GSSG) cycle is critical to paraquat resistance of tall fleabane. The decisive contribution of a functional GSH/GSSG cycle to paraquat resistance through an enhancement of GR activity in this weed was further confirmed by an experiment of exogenous application of ascorbate.