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It was reported that temperature affects the distribution of <i>Wolbachia</i> in the host, but only a few papers reported the effect of the interaction between high temperature and <i>Wolbachia</i> on the biological characteristic of the host. Here, we set four treatment <i>Drosophila melanogaster</i> groups: <i>Wolbachia</i>-infected flies in 25 °C (W⁺M), <i>Wolbachia</i>-infected flies in 31 °C (W⁺H), <i>Wolbachia</i>-uninfected flies in 25 °C (W^-M), <i>Wolbachia</i>-uninfected flies in 31 °C (W^-H), and detected the interaction effect of temperature and <i>Wolbachia</i> infection on the biological characteristic of <i>D. melanogaster</i> in F₁, F₂ and F₃ generations. We found that both temperature and <i>Wolbachia</i> infection had significant effects on the development and survival rate of <i>D. melanogaster</i>. High temperature and <i>Wolbachia</i> infection had interaction effect on hatching rate, developmental durations, emergence rate, body weight and body length of F₁, F₂ and F₃ flies, and the interaction effect also existed on oviposition amount of F₃ flies, and on pupation rate of F₂ and F₃ flies. High temperature stress reduced the <i>Wolbachia</i> vertical transmission efficiency between generations. These results indicated that high temperature stress and <i>Wolbachia</i> infection had negative effects on the morphological development of <i>D. melanogaster</i>.