Dynamic changes in global methylation and plant cell death mechanism in response to NiO nanoparticles

This report is a first comprehensive work on the potential of engineered nickel oxide nanoparticles affecting the epigenome and modulating global methylation leading to retention of transgenerational footprints. Nickel oxide nanoparticles (NiO-NPs) are known to instigate extensive phenotypic and physiological damage to plants. In the present work, it was shown that exposure to increasing concentrations of NiO-NP-induced cell death cascades in model systems, Allium cepa and tobacco BY-2 cells. NiO-NP also generated variation in global CpG methylation; its transgenerational transmission was shown in affected cells. Plant tissues exposed to NiO-NP showed progressive replacement of essential cations, like Fe and Mg, as seen in XANES and ICP-OES data, providing earliest signs of disturbed ionic homeostasis. Fluorescent staining based confocal microscopy confirmed upsurge of H₂O₂ and nitric oxide after NiO-NP exposure. A NiO-NP concentration gradient-based switching-on of the cell death cascades was observed when autophagosomes were seen in samples exposed to lower and median concentrations of NiO-NP (10-125 mg L^-1). The apoptotic cell death marker, caspase-3 like protein, was noted in the median to higher doses (50-500 mg L^-1), and leakage of lactate dehydrogenase marking necrotic cell death was observed in samples exposed to the highest doses (125-500 mg L^-1) of NiO-NP. Concomitant increase of DNA hypermethylation (quantified by ELISA-based assay) and genomic DNA damage (evaluated through Comet-based analyses) was recorded at higher doses of NiO-NP. MSAP profiles confirmed that global methylation changes incurring in the parental generation upon NiO-NP exposure were transmitted through the two subsequent generations of BY-2 cells which was supported by data from A. cepa, too. Thus, it was evident that NiO-NP exposure incited DNA hypermethylation, as an aftermath of oxidative burst, and led to induction of autophagy, apoptotic and necrotic cell death pathways. Global methylation changes induced by NiO-NP exposure can be transmitted through subsequent cell generations.