Abstract: Different origins Bt cotton cultivars, including DP410B (conventional cultivar) and Daiza No.1 (hybridized cultivar) from US and Sikang No.1 (conventional cultivar) and Sikang No.3 (hybridized cultivar) from China, were taken as the test materials to investigate the effects of high temperature (37 ℃) and different humidity (50%, 70%, and 90%) on the leaf Bt protein expression of Bt cotton. At high temperature, temperature and humidity had no significant effects on the leaf Bt protein expression of the cultivars at peak squaring stage. At peak flowering stage, as compared with the control (25-30 ℃ and 60%-70% humidity), 37 ℃ and 50% humidity decreased the leaf Bt protein content of conventional cultivars significantly by 2.6%-3.0%. At peak bolling stage, compared with the control, 37 ℃ and 50% humidity decreased the leaf Bt protein content of DP410B, Sikang No.1, and Sikang No.3 significantly by 3.3%-5.8%. Among the four cultivars, DP410B and Daiza No.1 had the highest leaf Bt protein content, while Sikang No.1 had the lowest one.