Diclofop-Methyl Increases the Proton Permeability of Isolated Oat-Root Tonoplast

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Abstract
Diclofop-methyl (methyl ester of 2-[4-(2'',4''-dichlorophenoxy)phenoxy]propionate; 100 micromolar) and diclofop (100 micromolar) inhibited both ATP- and PPi-dependent formation of H+ gradients by tonoplast vesicles isolated from oat (Avena sativa L., cv Dal) roots. Diclofop-methyl (1 miromolar) significantly reduced the steady-state H+ gradient generated in the presence of ATP. The ester (diclofop-methyl) was more inhibitory than the free acid (diclofop) at pH 7.4, but this relative activity was reversed at pH 5.7. Neither compound affected the rate of ATP or PPi hydrolysis by the proton-pumping enzymes. Diclofop-methyl (50, 100 micromolar), but not diclofop (100 micromolar), accelerated the decay of nonmetabolic H+ gradients established across vesicle membranes. Diclofop-methyl (100 micromolar) did not collapse K+ gradients across vesicle membranes. Both the (+)- and (-)-enantiomers of diclofop-methyl dissipated nonmetabolic H+ gradients established across vesicle membranes. Diclofop-methyl, but not diclofop (each 100 micromolar), accelerated the decay of H+ gradients imposed across liposomal membranes. These results show that diclofop-methyl causes a specific increase in the H+ permeability of tonoplast.