Carbon nanomaterials (CNMs) are novel engineered nanomaterials and have been used widely. Their toxic effects on terrestrial plants in soil matrix require careful investigation. In this study, white clover (Trifolium repens L.) was grown in a potted soil with graphene oxide (GO) at levels of 0.2%, 0.4% and 0.6% and the effects of GO on the growth and nutrient uptake of white clover were evaluated after 50 and 100 days of exposure. GO exposure showed adverse effects on seedling growth, photosynthetic parameters and nutrient uptake in shoots, and the effect was more significant with increasing concentration and exposure time. Compared with the control, GO at the highest level of 0.6% decreased plant height, leaf and stem dry weights, total chlorophyll content and net photosynthetic rate by 43.7%, 45.7%, 43.4%, 32% and 85.7%, respectively, after 100 d of exposure, and N, K, Cu, Zn, Fe, Mo, B, Si contents decreased by 19.5%, 20.1%, 12.6%, 25.0%, 12.9%, 26.0%, 18.9%, 23.0%, respectively. Furthermore, the electrolyte leakage, lipid peroxidation, reactive oxygen species, antioxidant enzyme activities were all increased by GO, especially at high dose and long exposure. These results indicate that GO can suppress plant growth by oxidative stress, photosynthesis inhibition, and nutrient imbalance. Display omitted •Data on the toxicity of graphene oxide (GO) to white clover in soil matrix were provided.•GO adversely affected plant growth and photosynthesis.•GO induced oxidative stress in white clover.•A decrease in nutrient levels was observed in white clover after GO exposure.