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The present study evaluated the effects of biofertilizers on qualitative and quantitative traits of <i>Nigella sativa</i> L. The experiment was conducted based on a randomized complete block design with 10 treatments and three replications. The treatments included control (no fertilization), U (100% chemical fertilizer as urea at 53.3 kg ha⁻¹, Nb (Biofertilizer, Azotobacter vinelandii), Pb (Biofertilizer, Pantoea agglomerans and Pseudomonas putida), Kb (Biofertilizer, Bacillus spp.), NPKb (NPK combined biofertilizers), Nb + 50% U, Pb + 50% U, Kb + 50% U, and NPKb + 50% U. The highest oil percentage was 46.33 percent related to NPK(b) + U50%, and the highest oil yield was 466.43 kg/ha related to NPK(b) + U50%. The highest seed yield (1006.64 kg/ha) was observed in the plants treated with NPK(b) + U50%. The treatment of K(b) + U50% increased myristic acid by 0.35%. The NPK(b) + U50% treatment reduced palmitic, stearic, and linolenic acid by 11.3, 2.01, and 0.31%, respectively. The highest oleic acid (27.85%) and linoleic acid (56.6%) were obtained from the treatment of NPK(b) + U50%, respectively. The plants treated with NPK(b) + U50% showed the highest seed N percentage (%), P content (mg g⁻¹) and K content (mg g⁻¹). In general, NPK(b) + U50% is the best treatment in line with sustainable agriculture.