Biochemical responses of the aquatic higher plant Lemna gibba to a mixture of copper and 1,2-dihydroxyanthraquinone: synergistic toxicity via reactive oxygen species.

Metals and polycyclic aromatic hydrocarbons (PAHs) are known to be toxic to plants. Because metals and PAHs often are cocontaminants in the environment, plants can be subjected to damage caused by their combined effects. We recently found that copper and an oxygenated PAH (1,2-dihydroxyanthraquinone [1,2-dhATQ]) synergistically are toxic to plants. This synergistic toxicity was linked indirectly to production of reactive oxygen species (ROS). In this study, plant growth, chlorophyll pigments, protein accumulation, and ROS production were chosen as endpoints to assess the mechanism of toxicity of copper and 1,2-dhATQ to Lemna gibba in more detail. Because copper and PAHs can generate ROS, we assayed for specific antioxidant enzymes: Superoxide dismutase (SOD), glutathione reductase (GR), and ascorbate peroxidase (APX). Copper treatment at a concentration that did not cause growth inhibition resulted in upregulation of Mn SOD, Cu-Zn SOD, and APX. At a level that moderately was toxic to plants, 1,2-dhATQ did not alter significantly the levels of these antioxidant enzymes. However, a synergistically toxic mixture of copper plus 1,2-dhATQ upregulated Cu-Zn SOD, Mn SOD, and GR, although APX activity was downregulated. When plants were treated with the ROS scavenger dimethyl thiourea (DMTU), enhanced toxicity and formation of ROS caused by the mixture both were diminished substantially. However, 1,2-dhATQ toxicity was not affected significantly by DMTU. Based on this study, the toxicity caused by the mixture of copper plus 1,2-dhATQ directly can be connected to elevated levels of ROS.

Similar stress responses are elicited by copper and ultraviolet radiation in the aquatic plant Lemna gibba: implication of reactive oxygen species as common signals.

Metals and ultraviolet (UV) radiation are two environmental stressors that can cause damage to plants. These two types of stressors often impact simultaneously on plants and both are known to promote reactive oxygen species (ROS) production. However, little information is available on the potential parallel stress responses elicited by metals and UV radiation. Using the aquatic plant Lemna gibba, we found that copper and simulated solar radiation (SSR, a light source containing photosynthetically active radiation (PAR) and UV radiation) induced similar responses in the plants. Both copper and SSR caused ROS formation. The ROS levels were higher when copper was combined with SSR than when applied with PAR. Higher concentrations of copper plus PAR caused toxicity as monitored by diminished growth and chlorophyll content. This toxicity was more pronounced when copper was combined with SSR. Because the generation of ROS was also higher when copper was combined with SSR, we attributed this enhanced toxicity to elevated levels of ROS. In comparison to PAR-grown plants, SSR treated plants exhibited elevated levels of superoxide dismutase (SOD) and glutathione reductase (GR). These enzyme levels were further elevated under both PAR and SSR when copper was added at concentrations that generated ROS. Interestingly, copper treatment in the absence of SSR (i.e. copper plus PAR) induced synthesis of the same flavonoids as those observed in SSR without copper. Finally, addition of either dimethyl thiourea or GSH (two common ROS scavengers) lowered in vivo ROS production, alleviated toxicity and diminished induction of GR as well as accumulation of UV absorbing compounds. Thus, the potential of ROS being a common signal for acclimation to stress by both copper and UV can be considered.