| Summary: | The common, broad-spectrum herbicide diuron poses some risks to the environment due to its long persistence and high toxicity. Therefore, the effective monitoring of diuron residues will inform efforts to assess its impacts on ecosystems. In this study, we evaluated the toxicity targets of diuron in the model aquatic macrophyte <i>Lemna minor</i> at the physiological (growth and photosynthetic efficiency), biochemical (pigment biosynthesis and reactive oxygen species (ROS) levels), and molecular (<i>rbcL</i> transcript) levels. The toxicity of diuron was detectable after 48 h of exposure and the order of sensitivity of toxicity endpoints was gene transcription > maximum electron transport rate (ETR<sub>max</sub>) > non-photochemical quenching (NPQ) > maximum quantum yield (<i>F<sub>v</sub>/F<sub>m</sub></i>) > ROS > fresh weight > chlorophyll <i>b</i> > chlorophyll <i>a</i> > total frond area > carotenoids. Under diuron stress, pigment, ROS, and gene transcript levels increased while frond area, fresh weight, and photosynthesis (<i>F<sub>v</sub>/F<sub>m</sub></i> and ETR<sub>max</sub>) gradually decreased with the increasing duration of exposure. Notably, ROS levels, <i>F<sub>v</sub>/F<sub>m</sub></i>, frond area, and fresh weight were highly correlated with diuron concentration. The growth endpoints (frond area and fresh weight) showed a strong negative correlation with ROS levels and a positive correlation with <i>F<sub>v</sub>/F<sub>m</sub></i> and ETR<sub>max</sub>. These findings shed light on the relative sensitivity of different endpoints for the assessment of diuron toxicity.
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