Water shortage is a global challenge and affects crop growth and development seriously. Stoma is the main channel of plant water transpiration. Water transpiration through stomata is an endothermic process and affects temperature of plants, especially leaf surface. In this study, we established a method for screening maize seedling mutant with abnormal leaf temperature by far infrared imaging. We found that seedling mutants with abnormal leaf temperature manifested different drought tolerance. Mutants with lower leaf temperature demonstrated faster moisture loss rate, poorer drought tolerance, higher osmotic potential, lower leaf relative water content, more accumulation of hydrogen peroxide, more serious cell membrane damage and more robust root systems and biomass under drought treatment, which are opposite in maize seedling mutants with higher leaf surface temperature compared with the corresponding control. Taken together, the method we established is an effective way to screen maize mutants with abnormal drought response.