Sorgoleone, the oxidized quinone form of a hydrophobic p-benzoquinone was first isolated from Sorghum root exudates. Sorgoleone is a potent inhibitor of growth in several annual weed species and causes tissue bleaching at concentrations of <25 μ M. These investigations were designed to determine if soreoleone's allelopathic activity was related to an inhibition of photosynthetic electron transport. The effect of sorgoleone versus DCMU (diuron) on inhibition of O2 evolution by broken wheat thylakoids, and in oxygenevolving PSII membranes containing QA and QB primary and secondary electron acceptors in PSII was determined. Sorgoleone was a potent inhibitor of O2 evolution in this system with ∼ 0.04 and 0.78 μ M concentrations required for 50 and 100% inhibition as compared to -0.11 and 2.0 μ M DCMU, respectively. Sorgoleone caused no significant inhibition of PSI mediated photooxidation of ascorbate/dichlorophenolindophenol, establishing that the locus of inhibition by sorgoleone was within the PSII complex. The effect of trypsin treatment of chloroplasts and PSII membranes on sensitivity to inhibition by DCMU and sorgoleone was examined. The comparison of DCMU and sorgoleone upon the formation and decay of flash-induced chlorophyll a variable fluorescence indicates that sorgoleone specifically inhibited the oxidation of QA by QB.