The effect of diuron [3-(3,4-dichlorophenyl)-1,1-dimethylurea], desmedipham [ethyl m-hydroxycarbanilate carbanilate(ester)], propanil (3′,4′-dichloropropionanilide), and dibromothymoquinone (DBMIB) (2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone) on proton translocation and photophosphorylation in isolated spinach (Spinacia oleracea L.) chloroplast fragments was investigated. In the absence of added cofactors, O2, or artificial electron acceptors, cyclic electron transport occurred, which was coupled to energy conservation. Under aerobic conditions O2 acted as the terminal acceptor in non-cyclic electron transport. Proton translocation and photophosphorylation in the cyclic process were enhanced by diuron, desmedipham, and propanil, while in the non-cyclic process they were inhibited by all three herbicides. DBMIB inhibited proton translocation and photophosphorylation in both processes. Proton translocation and its enhancement increased with increasing light intensities. The finding that the plastoquinone (PQ) antagonist DBMIB disrupted cyclic as well as noncyclic electron flow, while diuron enhanced the cyclic and inhibited the noncyclic process, indicated that the acceptor site for endogenously-cycling electrons must lie between the active site of diuron inhibition and PQ, The close similarity in the behavior of diuron, desmedipham, and propanil suggests that their site of action is the same.