Strontium marking technology is commonly used for extensive marking in stock enhancement and releasing activities. In order to explore the feasibility of this technology for juvenile Takifugu obscurus, five different concentrations of strontium (0, 12, 18, 36, and 72 mg/L) were set up by strontium chloride hexahydrate (SrCl2·6H2O) and T. obscurus were immersed for 7 days. Then, T. obscurus were reared in non-additive water for 25 days. The results showed that the survival rate of all groups was 100%, except the 72 mg/L group, after 7 days of immersion. Moreover, the growths of all the marked groups were better than that of the control group. There was no significant difference between the control and marked groups, except for the 18 mg/L group, indicating that the appropriate concentration of strontium might have positive effect for T. obscurus. The strontium to calcium (Sr/Ca) ratios of otoliths in the marked groups increased with increasing concentration and time, which were higher than the baseline, respectively. Then, the Sr/Ca ratios returned to the original level, but the peak value was still retained, suggesting that the success rate of the strontium marking reached 100%. Notably, the residual strontium content of 18 mg/L group was insufficient for causing negative effects for T. obscurus after 25 days. Therefore, in consideration of the growth, survival, and effect of strontium marking on otolith, 18 mg/L is an appropriate concentration for strontium marking of juvenile T. obscurus. This study confirmed the feasibility of strontium marking for T. obscurus and provided a new approach to rationally and scientifically evaluate the stock enhancement and releasing efficiency of T. obscurus.