Detoxifying processes during kanamycin-induced stress to Arabidopsis thaliana seedling growth

Abstract

The increasing antibiotic application in agriculture has raised a potential risk to human health worldwide. Although high concentrations of antibiotics are lethal for plants without a resistant gene, it is not clear what detoxifying processes act in plants under low antibiotic concentration. Arabidopsis thaliana was selected to characterize the effects of kanamycin antibiotic toxicity on seedling growth in MS plates and pots, respectively. We investigated only control plants treated with kanamycin; but without comparison with transgenic ones, since we had no positive control transgenic plants. Gene expression profiles from plant material in pots were evaluated by customized oligonucleotide microarray. Morphological investigation indicated that kanamycin concentration is negatively correlated with A. thaliana seedling growth. Gene expression profile analysis detected that 542 and 116 genes were up-and down-regulated over five-fold in leaf tissue under the influence of kanamycin. The glutathione S-transferase and peroxidase gene families were involved in antioxidant defense mechanisms by preventing the accumulation of toxic products of reactive oxygen species (ROS). An exceeding amount of ROS after antibiotic-induced oxidative burst stimulated several over-presented processes for the balance between ROS and antioxidants. The significant increase in detoxification enzymes may indicate that efficient antioxidant machinery is the main method to ensure survival.

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“…However, the selected plants can be affected by their resistance properties. High concentrations caused seedling etiolation and plant death ( Chen et al, 2020 ). A concentration of 50 ppm kanamycin was effectively used in the selection of transformants of Glycine max L. ( Isda, 2012 ) and Saccharum officinarum ( Fibriani et al, 2019 ).…”

“…Moreover, each plant species has a different tolerance level for various concentrations of kanamycin antibiotics. Seedlings of wild-type A. thaliana tolerated up to 1,400 ppm kanamycin ( Chen et al, 2020 ), Gossypium hirsutum L. tolerated up to < 50 ppm kanamycin ( Unbeck et al, 1989 ), Lycopersicon esculentum Mill was putatively transformed under 100 ppm kanamycin ( Subaila and Saleh, 2010 ), and Artemisia annua L. tolerated up to < 20 ppm kanamycin ( Chen et al, 2000 ).…”

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“…A similar observation was reported by Wang et al [17] at a high concentration of kanamycin. Chen et al [48] reported the observation of leaf yellowing in Arabidopsis explants cultured on kanamycin-containing media. According to Wang et al (2015) [17], the yellowing and subsequent death of cells in green organs are caused by an interference of kanamycin with protein synthesis in the chloroplasts and mitochondria of plant cells that do not possess the detoxifying ability of the kanamycin resistance gene nptII.…”