Oxidative response of rice (Oryza sativa L.) seedlings to quinolone antibiotics and its correlation with phyllosphere microbes and antibiotic resistance genes

Abstract

With the increasing use of veterinary antibiotics, quinolone antibiotics may enter farmland systems after livestock manure has been composted. However, the phytotoxicity mechanism of antibiotics in crops is still unclear. In this study, the oxidative responses of rice (Oryza sativa L.) seedlings to three typical quinolone antibiotics and their underlying mechanisms were investigated. The bioconcentration factor values were 1.47, 0.55, and 0.23 in the levofloxacin, enrofloxacin and norfloxacin treatment, respectively. The inhibitory effects on rice seedlings were in the order of levofloxacin > enrofloxacin > norfloxacin, which may be due to the high uptake of levofloxacin. The H₂O₂ level, MDA content, and ion leakage rate increased significantly (p < 0.05), and cell plasmolysis was observed, showing that antibiotics can cause membrane lipid peroxidation and damage the cell membrane structure. Antioxidant enzyme activities (superoxide dismutase, catalase, and peroxidase) changed with the antibiotic concentration. Integrated biomarker response analysis showed that levofloxacin caused the greatest oxidative stress in rice seedlings. Transcriptomic analysis identified 5880 differentially expressed genes, and these were annotated as 20 biological functions; the greatest abundances were cellular and metabolic processes, cell part, and membrane part and organelle; SOD and CAT related genes were up-regulated. The richness and diversity of the phyllosphere microbial community decreased significantly (p < 0.05) and the microbiome changed at the phylum and genus levels. The H₂O₂ level was correlated with changes in phyllosphere microbial communities. The number of antibiotic resistance genes (ARGs) and mobile genetic elements decreased, while their abundance increased. In conclusion, enrofloxacin exposure not only affects the microbial community but may also affect the ARGs carried by microbes. The relative abundance of MGEs and ARGs was significantly positively correlated (R² = 0.760, p = 0.0148), indicating that MGEs can significantly promote the spread of ARGs.

Keywords: Antibiotic resistance genes; Bioconcentration factor; Integrated biomarker response analysis; Phyllosphere microbes; Quinolone antibiotics; Transcriptomic analysis.