Localization of electron transport inhibition in plastoquinone reactions

Reduction kinetics of P700 following a short flash are measured in spinach chloroplasts after oxidation of the electron carriers between the two photoreactions by far-red light. Three features of the kinetics allow us to localize simultaneously inhibition at different sites between photoreaction II and the reducing site of plastoquinol. These are the initial lag, the halftime and the area under the transient of the P700 absorbance change, which indicate the electron transfer time from photoreaction II to the reducing site of plastoquinol, the rate of plastoquinol oxidation, and the number of electrons transferred to the special plastoquinone B functioning as secondary electron acceptor of photosystem II, respectively. As an additional diagnostic parameter for inhibition before and after the plastoquinone pool, the area under the transient of the P700 absorbance change is used after long flashes. This area is proportional to the amount of reduced plastoquinone as shown by the absorbance change at 265 nm. The effects of 3-(3',4'-dichlorophenyl)-1,1-dimethylurea (DCMU) and 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB) are compared with those of 2-bromo-4-nitrothymol, 2,4-dinitrophenyl ether of 2-iodo-4-nitrothymol, and Illoxan as representatives for new classes of inhibitors. While 2-halogeno-4-nitrothymols inhibit the reduction of plastoquinone similarly to DCMU, their diphenyl ether derivatives inhibit selectively the oxidation of plastoquinol.