ABSTRACT
Selenium (Se) is an essential element for humans and animals that is required for key antioxidant reactions, but can be toxic at high concentrations. We have investigated the effect of Se in the form of selenite on coffee cell suspension cultures over a 12-day period. The antioxidant defence systems were induced in coffee cells grown in the presence of 0.05 and 0.5 mm sodium selenite (Na2SeO3). Lipid peroxidation and alterations in antioxidant enzymes were the main responses observed, including a severe reduction in ascorbate peroxidase activity, even at 0.05 mm sodium selenite. Ten superoxide dismutase (SOD) isoenzymes were detected and the two major Mn-SOD isoenzymes (bands V and VI) responded more to 0.05 mm selenite. SOD band V exhibited a general decrease in activity after 12 h of treatment with 0.05 mm selenite, whereas band VI exhibited the opposite behavior and increased in activity. An extra isoenzyme of glutathione reductase (GR) was induced in the presence of selenite, which confirmed our previous results obtained with Cd and Ni indicating that this GR isoenzyme may have the potential to be a marker for oxidative stress in coffee.
ABSTRACT
The antioxidant responses of coffee (Coffea arabica L.) cell suspension cultures to cadmium (Cd) were investigated. Cd accumulated very rapidly in the cells and this accumulation was directly correlated with an increase in applied CdCl(2) concentration in the external medium. At 0.05mM CdCl(2), growth was stimulated, but at 0.5mM CdCl(2), the growth rate was reduced. An alteration in activated oxygen metabolism was detected by visual analysis as well as by an increase in lipid peroxidation at the higher CdCl(2) concentration. Catalase (CAT; EC 1.11.1.6), glutathione reductase (GR; EC 1.6.4.2) and superoxide dismutase (SOD; EC 1.15.1.1) activity increased, particularly at the higher concentration of CdCl(2). Ascorbate peroxidase (APX; EC 1.11.1.11) activity was increased at the lower CdCl(2) concentration used, but could not be detected in cells growing in the higher CdCl(2) concentration after 24h of growth, whilst guaiacol peroxidase (GOPX; EC 1.11.1.7) did not show a clear response to Cd treatment. An analysis by non-denaturing PAGE followed by staining for enzyme activity, revealed one CAT isoenzyme, nine SOD isoenzymes and four GR isoenzymes. The SOD isoenzymes were differently affected by CdCl(2) treatment and one GR isoenzyme was shown to specifically respond to CdCl(2). The results suggest that the higher concentrations of CdCl(2) may lead to oxidative stress. The main response appears to be via the induction of SOD and CAT activities for the removal of reactive oxygen species (ROS), and by the induction of GR to ensure the availability of reduced glutathione for the synthesis of Cd-binding peptides, which may also be related to the inhibition of APX activity probably due to glutathione and ascorbate depletion.
