The nitrate uptake and growth of wheat were more inhibited under single-layer graphene oxide stress compared to multi-layer graphene oxide

Although the phytotoxicity of graphene-based materials has been investigated extensively, the effects of different graphene-based materials on nutrient uptake in plants remain unclear. Here, we analyzed the differences in phytotoxicity between single-layer graphene oxide (sGO) and multi-layer graphene oxide (mGO) by analyzing the growth status and nitrate (NO3-) accumulation in wheat plants at 0, 100, 200, 400, and 800 mg L-1 graphene oxide supply. Both sGO and mGO displayed concentration-dependent inhibitory effects on biomass, root length, number of lateral roots, and nitrogen (N) nutrient status. Treatment with 400 mg L-1 sGO caused 0.9-, 1.3-, and 1-fold higher reductions in NO3--N, assimilated N, and total N concentrations in roots, respectively, than mGO treatment. Analysis of root oxidative stress and in situ NO3- uptake revealed that sGO caused more significant damage to the root tip and a lower NO3- net influx rate than mGO. In addition, the expression of NO3- transporter (NRT) genes in roots, including NRT1.5, NRT2.1, NRT2.2, NRT2.3, and NRT2.4, under sGO treatment were lower than those under mGO treatment. Overall, sGO treatment induced a more severe inhibitory effect on root growth and NO3- uptake and accumulation than mGO treatment, accompanied by significant suppression of the expression of NRTs in sGO-treated roots. This study provides a physiological and molecular basis for studying the phytotoxic effects of various sizes of graphene oxide.