Enzymatic and Non-enzymatic Antioxidant Responses of Alfalfa Leaves and Roots Under Different Salinity Levels

The effect of increasing NaCl concentrations on biomass, hydrogen peroxide (H2O2), ascorbic acid (ASC), proline and total thiol, and the activity of some antioxidant enzymes in alfalfa (Medicago sativa L. cv. Gara-Yonjeh) were investigated. The dry weights of roots and shoots with increasing NaCl concentrations decreased progressively, and the strongest toxicity was detected at NaCl treatment of 200 mM. Superoxide dismutase (SOD) activity in the leaves increased gradually up to NaCl concentrations of 100, while the higher concentration of NaCl reduced SOD activity in both leaves and roots. The maximum levels of ascorbate peroxidase (APX) activity were increased at 150 mM and 100 mM NaCl in leaves and roots of Gara-Yonjeh, respectively. Peroxidase (POD) activity in roots of Gara-Yonjeh increased (82% at 200 mM) by salinity, while it decreased (43% at 200 mM) in leaves. In contrast, catalase (CAT ) activitiy increased (84% at 200 mM) in leaves, and decreased (57% at 200 mM) in the roots of Gara-Yonjeh. Electrophoresis analysis suggested that different patterns in SOD, CAT and POD isoenzymes depend on NaCl concentrations, and the staining intensities of these isoforms are supported the results obtained from the spectrophotometric determinations. In POD and CAT, activity of isoform III was detected at all concentrations, by a “low-high-low” pattern, with the maximum activity at 50 mM of NaCl. Results imply that the function of antioxidant systems in higher NaCl concentration is responsible for the salt tolerance observed in Gara-Yonjeh.