Stress response to oxytetracycline and microplastic-polyethylene in wheat (Triticum aestivum L.) during seed germination and seedling growth stages

Much efforts have been devoted to clarify the phytotoxicity of individual contaminants in plants, such as individual antibiotic and microplastic; however, little is known about the phytotoxicity of their combined exposure. Here, we investigated the effects of individual and combined exposure of wheat (Triticum aestivum L.) (Xiaoyan 22) to oxytetracycline (OTC) and polyethylene (PE) microplastics using physiological and metabolic profilings. During the seed germination stage, OTC induced phytotoxicity, as observed through the changes of root elongation, sprout length, fresh weight and the vitality index, with significant effect at the 50 and 150 mg·L-1 levels; the effect of PE microplastics depended on the OTC level in the combined exposure groups. During seedling cultivation, catalase (CAT) and ascorbate peroxidase (APX), as antioxidant enzyme indices, were sensitive to OTC exposure stress, although OTC was not determined in leaves. Untargeted metabolomics of wheat leaves revealed OTC concentration-, metabolite class- and PE-dependent metabolic responses. Dominant metabolites included carboxylic acids, alcohols, and amines in the control group and all treatment groups. Compared to only OTC treatment, PE reprogrammed carboxylic acid and alcohol profiles in combined exposure groups with obvious separation in PLS-DA. Combined exposure induced fewer metabolites than OTC exposure alone at the 5 and 50 mg·L-1 levels. The shared metabolite numbers were higher in the OTC groups than in the PE-OTC groups. Pathway enrichment analysis showed a drift in metabolic pathways between individual and combined exposure to OTC and PE, which included glyoxylate and dicarboxylate metabolism, amino acid metabolism and isoquinoline alkaloid biosynthesis. Among metabolites, aromatic acids and amino acids were more sensitive to combined exposure than individual exposure. These results contribute to clarifying the underlying mechanisms of phytotoxicity of individual and combined exposure to OTC and PE.