Abstract: The natural reproduction of fish is a physiological response process under the stimulation of specific environment. The environment condition of fish spawning habitat is very important for the reproduction of fish. Currently, there is no quantitative study on the demand for natural reproduction of Gymnocypris przewalskii. In combination with the in situ investigation of the natural spawning grounds of G. przewalskii, artificial spawning environment was simulated in the laboratory to answer the environmental requirement for the natural reproduction of G. przewalskii. The results showed that the occurrence of spawning activities of G. przewalskii was closely related to the water temperature, water depth, flow rate, light and substrate factors. The results showed that the natural mating induction rate and reproductive effect can be significantly increased in the substrate of pebble than that in the sand or the mud substrate. The suitable range of the water depth, water velocity, water temperature for spawning is 0.15—0.2 m, 0.2—0.4 m/s, and 10.8—14.3℃, respectively. Increasing the everyday’s dark cycle is beneficial for the occurrence of spawning. Under the controlled environment, when the water depth is above 0.45 m, water velocity is more than0.8 m/s, and the water temperature is higher than 17℃ or lower than 6℃, there will be no spawning activities, which maybe the limit conditions required for the natural reproductive.This study elucidated the environmental parameters for the spontaneous spawning and reproduction of G. przewalskii in a controlled environment, and successfully constructs a simulated spawning environment. It can provide technical supports and new ideas in the natural spawning habitat investigation and contribute for improving the artificial breeding technologies of G. przewalskii.