SYNERGISTIC EFFECTS OF A PHOTOOXIDIZED POLYCYCLIC AROMATIC HYDROCARBON AND COPPER ON PHOTOSYNTHESIS AND PLANT GROWTH: EVIDENCE THAT IN VIVO FORMATION OF REACTIVE OXYGEN SPECIES IS A MECHANISM OF COPPER TOXICITY

Heavy metals and polycyclic aromatic hydrocarbons (PAHs) are often cocontaminants in industrialized environments, yet little is known about either the extent or mechanisms of their cotoxicity. To address this shortfall, the combined effects of an oxygenated PAH, 1,2-dihydroxyanthraquinone (1,2-dhATQ), and a heavy metal, Cu super(2+), on photosynthesis and growth of the duckweed (Lemna gibba) were evaluated. Using assays of chlorophyll a fluorescence and photosystem I activity, 1,2-dhATQ inhibited electron transport at the cytochrome b sub(6)/f complex. Conversely, Cu super(2+) alone (at low concentrations) had little effect on photosynthesis. When Cu super(2+) was combined with 1,2-dhATQ, an increase in transient and steady-state chlorophyll a fluorescence quenching occurred relative to 1,2-dhATQ alone. Treatment of isolated thylakoid membranes with 1,2-dhATQ inhibited whole-chain linear electron transport, measured as O sub(2) consumption using methyl viologen as the electron acceptor. However, Cu super(2+) plus 1,2-dhATQ resulted in active O sub(2) consumption with or without methyl viologen as an electron acceptor. From these data, we conclude that 1,2-dhATQ renders the plastoquinone pool to a highly reduced state by inhibiting at cytochrome b sub(6)/f. Then, Cu super(2+) is able to mediate the transfer of electrons from reduced plastoquinone to O sub(2), forming reactive oxygen species. At the whole-organism level, when Cu super(2+) and 1,2-dhATQ were mixed at concentrations that resulted in the above-mentioned impacts on photosynthesis, synergistic inhibition of plant growth was observed. This suggests a catalytic mechanism of toxicity for redox active metals, a process that could be instrumental in explaining their impacts at low concentrations.