In a field trial, the effect of soaking three cultivars of mustard (Brassica junceaL. Czern &Coss.) namely ‘RK‐8203’,’PR‐18’, and ‘Varuna’ for 4 h in 0.0125, 0.025, 0.05, and 0.10% aqueous pyridoxine solution was studied on growth and yield characteristics. In addition to pyridoxine treatment, two controls (unsoaked and water‐soaked) were also maintained for comparison. The parameters studied included root length, leaf number, dry weight/plant, and leaf N, P, and K content at 70 days after sowing (DAS). Pods/plant, seeds/pod, 1000 seed weight, oil content, seed yield, and oil yield were measured at harvest. The pyridoxine requirement for optimum performance of ‘RK‐8203 ‘ was 0.05% and for ‘Varuna’ it was 0.0125%, whereas ‘PR‐18’ showed inhibitory effect to pyridoxine treatment. This variation in pyridoxine requirement of mustard cultivars for optimum performance seems to be the result of the difference in the native pyridoxine content in the seed. The results suggest that the performance of cultivarsof Brassica junceahaving low seed pyridoxine content could be improved by soaking the seeds in aqueous pyridoxine solution so that pyridoxine in seeds reaches the optimum level.