Arsenic (As) contamination has emerged as a serious public health concern worldwide because of its accumulation and mobility through the food chain. Therefore, the current study was planned to check the effect of Bacillus subtilis-synthesized iron oxide nano particles (Fe3O4 NP) on rice (Oryza Sativa L.) growth against arsenic stress (0, 5, 10 and 15 ppm). Iron oxide nanoparticles were extracellular synthesized from Bacillus subtilis with a desired shape and size. The formations of nanoparticles were differentiated through UV-Visible Spectroscopy, FTIR, XRD and SEM. The UV-Visible spectroscopy of Bacillus subtilis-synthesized nanoparticles showed that the iron oxide surface plasmon band occurs at 268 nm. FTIR results revealed that different functional groups (aldehyde, alkene, alcohol and phenol) were present on the surface of nanoparticles. The SEM image showed that particles were spherical in shape with an average size of 67.28 nm. Arsenic toxicity was observed in seed germination and young seedling stage. The arsenic application significantly reduced seed germination (35%), root and shoots length (1.25 and 2.00 cm), shoot/root ratio (0.289), fresh root and shoots weight (0.205 and 0.260 g), dry root and shoots weight (6.55 and 6.75 g), dry matter percentage of shoot (12.67) and root (14.91) as compared to control. Bacillus subtilis-synthesized Fe3O4 NPs treatments (5 ppm) remarkably increased the germination (65%), root and shoot length (2 and 3.45 cm), shoot/root ratio (1.24) fresh root and shoot weight (0.335 and 0.275 mg), dry root and shoot weight (11.75 and 10.6 mg) and dry matter percentage of shoot (10.40) and root (18.37). Results revealed that the application of Fe3O4 NPs alleviated the arsenic stress and enhanced the plant growth. This study suggests that Bacillus subtilus-synthesized iron oxide nanoparticles can be used as nano-adsorbents in reducing arsenic toxicity in rice plants.