Nitrate Reductase-Dependent Nitric Oxide Production Is Involved in Microcystin-LR-Induced Oxidative Stress in Brassica rapa

Histochemical and biochemical approaches were used to investigate the phytotoxicity induced by microcystin-LR (MC-LR) in the shoots of Brassica rapa seedlings. MC-LR exposure was able to induce oxidative stress by triggering the over-generation of reactive oxygen species (ROS) including superoxide anion radical (O-2 (au -) ) and hydrogen peroxide (H2O2) in the shoots of B. rapa. MC-LR exposure led to the significant increase in the concentration of endogenous nitric oxide (NO) in B. rapa. However, such increase was completely suppressed by the treatment with nitrate reductase (NR) inhibitor NaN3, while l-NMMA, a NO synthase (NOS) inhibitor, had only slight effect on the content of endogenous NO in MC-LR-treated plant. These data suggested that NR-dependent pathway was the main source for endogenous NO generation under MC-LR stress. Afterwards, treatment with NaN3 reduced the ROS generation, lipid peroxidation, and loss of membrane integrity in MC-LR-treated plant. MC-LR stress induced the increase in the expression of superoxide dismutase, ascorbate peroxidase, and catalase. However, such an effect could be reversed by the treatment with NaN3. These results indicate that NR-dependent NO production mediates MC-LR-induced oxidative stress by triggering the over-generation of ROS in B. rapa.