Methylene blue is widely used in various industrial branches. Due to insufficient treatment, its occurrence in wastewater is frequently detected, which may result in serious environment problems to aquatic organisms. Hydroponic experiments were conducted with rice seedlings (Oryza sativa L. cv. XZX 45) exposed to methylene blue to determine the effective concentration using relative growth rate and water use efficiency as response endpoints. Results showed that acute toxicity of methylene blue to rice seedlings was evident. Although a linear decrease in relative growth rate and water use efficiency was observed in rice seedlings with increasing methylene blue concentrations, relative growth rate of rice seedlings was more sensitive to change of methylene blue than water use efficiency. Using non-linear regression, EC-48 h values for 10%, 20% and 50% inhibition of the relative growth rate were estimated to be 1.54, 3.22 and 10.13 mg MB/L for rice seedlings exposed to methylene blue, respectively, while smaller EC were obtained for 96 h exposure. In conclusion, the toxic response of young rice seedlings to methylene blue is obvious and inhibitory effects are highly dependent on response endpoints and the duration of exposure period.