The herbicide Mesotrione herbicides are very effective in the control of a wide range of weeds that infest corn (Zea mays) fields. However, the biochemical and molecular bases of corn seedling tolerance to this herbicide have not been established so far. To understand the mechanisms of mesotrione detoxification in corn plants, the activities of the main primary proton (H+ ion) transport systems of the vacuolar and plasma membranes (H+-ATPases V- and P-types, and H+-PPase) of the cells from different tissues were analyzed, after post-emergence herbicide application. Thus, cell fractionation procedures on root, leaf and mesocotyl tissues were performed using differential membrane vesicle centrifugation and purification in sucrose density gradient. Hydrolytic activities of the proton pumps were measured by using a colorimetric method for phosphate released through enzymatic hydrolysis of the substrates adenosine-5'-triphosphate (ATP) and pyrophosphate (PPi). Photosynthetic parameters were analyzed as physiological markers of the different stages of plant detoxification. Such analysis demonstrated that, three days after herbicide application (DAA), mesotrione induced a reduction in the photosynthetic rate and Fv/Fm ratio, but no significant effect could be found after the fifth DAA. These data suggest that the treatment with mesotrione promoted a spatial and temporal regulation of the H+ pump activities. In all the root, leaf and mesocotyl tisues analyzed, at three DAA, a strong stimulation of the vacuolar H+-PPase activity was observed, varying from 100% to 600%. This activation was significantly reduced at the seventh DAA, but remained at least twice higher than the controls. On the other hand, the vacuolar and plasma membranes H+-ATPases were much less modulated by the herbicide treatment, exhibiting stimulations and inhibitions which did not change by more than 20 to 60% in relation to the activities found in membrane vesicles derived from untreated plants (controls). The results demonstrate that mesotrione can promote a differential activation of the main primary proton transport systems, suggesting that these ion-pumps are transporting enzymes essential for the detoxification-related processes in corn plants, likely by energizing compartmentalization of the herbicide molecules into the vacuoles or cellular excretion through the plasma membrane.
Zea mays; Herbicide; Phytotoxicity; H+-ATPase; V-ATPase; H+-PPase
