[Objectives]  Aphis gossypii Glover is a major pest of wolfberry crops. Chemical control is currently one of the main methods used to control the crop damage caused by this pest. The effectiveness of pyrethroids is generally thought to decrease with temperature increasing. [Methods]  The effect of temperature on the toxicity of pyrethroids to A. gossypii was investigated. The residual film method was used to determine the toxicity of three pyrethroids to A. gossypii. at 17, 27 and 37℃. An ATPase kit was used to measure the effect of the above three pyrethroids on mitochondrial Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase activity at different temperatures. Fluorescence polarization was used to study mitochondrial membrane fluidity in A. gossypii with DPH as a fluorescent probe. [Results]  In the three pyrethroids tested, the toxicity of deltamethrin increased, whereas that of phenvalerate and cyfluthrin, decreased with temperature increasing. At a pyrethroid concentration of 1×10^－4 mol /L, the inhibitory effect of all the three pyrethroids on Na⁺-K⁺-ATPase and Ca²⁺-Mg²⁺-ATPase activity decreased with temperature increasing. There was similar trend in the effect of the three pyrethroids on mitochondrial membrane fluidity at the three tested temperatures. Changes in DPH polarization caused by the three pyrethroids were greater at lower than higher temperatures, which is consistent with the generally accepted negative correlation between pyrethroid toxicity and temperature. [Conclusion]  The relationship between temperature and pyrethroid toxicity is complex, and ATPase and mitochondrial membrane fluidity play an important role in the general negative temperature coefficient of pyrethroid toxicity.