Assessment of the effects of metribuzin, glyphosate, and their mixtures on the metabolism of the model plant Lemna minor L. applying metabolomics.

Chemical plant protection products (PPPs) is a major group of xenobiotics that are being released in the environment. Although the effects of individual active ingredients (a.i.) on organisms have been studied, information on those of mixtures, is fragmented. Aquatic environments are being polluted by PPPs, posing serious risks for the environment, human, and other organisms. Based on the potential of the model aquatic plant Lemna minor L. in the assessment of PPPs-caused stresses, we have undertaken the task of developing a metabolomics approach for the study of the effects of metribuzin and glyphosate, and their mixtures. Bioassays revealed that metribuzin exhibit higher toxicity than glyphosate and metabolomics highlighted corresponding changes in its metabolome. Treatments had a substantial impact on plants' amino acid pool, resulting in elevated levels of the majority of the identified amino acids. Results indicate that the increased proteolytic activity is a common effect of the a.i. and their mixtures. Additionally, the activation of salicylate-signaling pathways was recorded as a response to the toxicity caused by mixtures. Among the identified metabolites that were discovered as biomarkers were γ-aminobutyric acid (GABA), salicylate, caffeate, α,α-trehalose, and squalene, which play multiple roles in plants' metabolism such as, signaling, antioxidant, and structure protection. No reports exist on the combined effects of PPPs on Lemna and results confirm the applicability of Lemna metabolomics in the study of the combined effects of herbicides and its potential in the monitoring of the environmental health of aquatic environments based on fluctuations of the plant's metabolism.