Diazotrophic rhizobacteria, trigger and enhance plant growth as well as yield through various mechanisms, so their use can reduce the application frequency of chemical fertilizers. Indole-3-acetic acid (IAA), a most common natural auxin influences several physiological processes of the plant's health. The present study is aimed to optimize the conditions for IAA production, along with assay for plant growth promoting traits of Bacillus subtilis DR2 (KP455653), which is a diazotrophic Gram positive, rod bacterium, isolated from rhizosphere of road side weed, Eragrostis cynosuroides from Danapur, Patna, Bihar, India. The screening for IAA production was done in JNFbˉ broth with tryptophan (1 g.l-1) and without tryptophan at pH 5.8, 30±2 °C temperature and 48 h incubation. 137.81 µg.ml-1 and 100.26 µg.ml-1 IAA was produced in Trp + and Trpmedia, respectively. Under various optimized conditions, maximum IAA was produced at 96 h incubation (137.81 µg.ml-1), 35 °C temperature (141.92 µg.ml-1), pH 7 (158.79 µg.ml-1), mannitol as carbon (160.85 µg.ml-1) and ammonium sulfate as nitrogen (162.93 µg.ml-1) sources with tryptophan at final concentration of 1.2 µg.ml-1 (168.09 µg.ml-1), which enhanced the production by 1.2 fold. The findings suggest that B. subtilis DR2 is a potent organism to be used as biofertilizer.