The effect of zinc oxide nanoparticles (ZnONPs) was studied in wheat (Triticum aestivum L.) seedlings under in vitro exposure conditions. To avoid precipitation of nanoparticles, the seedlings were grown in half strength semisolid Murashige and Skoog medium containing 0, 50, 100, 200, 400 and 500 mg L -1 of ZnONPs. Analysis of zinc (Zn) content showed significant increase in roots. In vivo detection using fluorescent probe Zynpyr-1 indicated accumulation of Zn in primary and lateral root tips. All concentrations of ZnONPs significantly reduced root growth. However, significant decrease in shoot growth was observed only after exposure to 400 and 500 mg L -1 of ZnONPs. The reactive oxygen species and lipid peroxidation levels significantly increased in roots. Significant increase in cell-wall bound peroxidase activity was observed after exposure to 500 mg L -1 of ZnONPs. Histochemical staining with phloroglucinol-HCl showed lignification of root cells upon exposure to 500 mg L -1 of ZnONPs. Treatment with propidium iodide indicated loss of cell viability in root tips of wheat seedlings. These results suggest that redox imbalances, lignification and cell death has resulted in reduction of root growth in wheat seedlings exposed to ZnONPs nanoparticles.