The propagation of strelitzia plants can be carried out in vitro as an alternative to combine the aseptic conditions of the culture medium with the use of strategies to promote germination and controlled abiotic conditions. However, this technique is still limited by the prolonged time and low percentage of seed germination, which is the most viable explant source, due to dormancy. Thus, the objective of this study was to evaluate the influence of chemical and physical scarification processes of seeds combined with gibberellic acid (GA3), as well as the effect of graphene oxide in the in vitro cultivation of strelitzia plants. Seeds were subjected to chemical scarification with sulfuric acid for different periods (10 to 60 min) and physical scarification (sandpaper), in addition to a control treatment without scarification. After disinfection, the seeds were inoculated in MS (Murashige and Skoog) medium with 30 g L−1sucrose, 0.4 g L−1PVPP (polyvinylpyrrolidone), 2.5 g L−1Phytagel®, and GA3at different concentrations. Growth data and antioxidant system responses were measured from the formed seedlings. In another experiment, the seeds were cultivated in vitro in the presence of graphene oxide at different concentrations. The results showed that the highest germination was observed in seeds scarified with sulfuric acid for 30 and 40 min, regardless of the addition of GA3. After 60 days of in vitro cultivation, physical scarification and scarification time with sulfuric acid promoted greater shoot and root length. The highest seedling survival was observed when the seeds were immersed for 30 min (86.66%) and 40 min (80%) in sulfuric acid without GA3. The concentration of 50 mg L−1graphene oxide favored rhizome growth, while the concentration of 100 mg L−1favored shoot growth. Regarding the biochemical data, the different concentrations did not influence MDA (Malondialdehyde) levels, but caused fluctuations in antioxidant enzyme activities.