Rice (Oryza sativa L.) is a staple food crop; most of it is consumed in nations where malnutrition is a serious problem, and its enrichment through biofortification can be used to efficiently combat hidden hunger. Here, we studied the effect of two zinc forms, i.e., zinc oxide nanoparticles (ZnO NPs) and sulfate salt (ZnSO4), at four different concentrations during the grain development period (after anthesis and continued once a week for up to 5 weeks) of the rice plant. During the rice growing season 2021–2022, all the experiments were conducted in a greenhouse (temperature: day 30 °C; night 20 °C; relative humidity: 70%; light period: 16 h/8 h, day/night). The main aim was to identify the effects of ZnO NPs on physical growth, biochemical parameters, nutrient acquisition, and crop yield. We have also highlighted the effects of NPs on zinc biofortification, and the end results illustrated that both zinc forms are capable of increasing grain yield. However, we found that even at low concentrations, ZnO NPs showed a significant increase in growth yield, whereas bulk did not show eminent results even at higher concentrations. Spikelet number per panicle was more than 50% and 38% in the case of ZnO NPs and ZnSO4, respectively. Similarly, stimulation in plant height was 25% with NPs treatment and only 3% with bulk treatment. The increase in grain per spike was 19% with ZnO NPs as compared to the control. Total chlorophyll, soluble sugar, amylose, and soluble protein contents were enhanced under ZnO NP treatment, which plays an excellent role in the regulation of various transcriptional pathways related to biofortification. We identified that foliar application at the flowering stage is more effective in comparison to the basal and tillering stages of the rice life cycle. ZnO NPs increased zinc content in rice grain by 55% as compared to traditional fertilization (~ 35%), with no adverse effects on human health. This study highlights that ZnO NPs could be used to increase zinc efficiency and as a safe fertilizer in the rice harvesting ecosystem.