The responses of soybean genotypes to high temperature for intrinsic tolerance was studied using Temperature Induction Response (TIR) technique in order identify the genotypes tolerant to high temperature stress. Seven soybean genotypes subjected to lethal and sub lethal temperatures showed significant variation for acquired thermotolerance. Thermotolerant genotypes ADT 1 and CoSoy1 identified by the TIR technique, demonstrated higher survival percentage, and lower growth reduction. Further the tolerant genotypes identified based on TIR also showed higher antioxidant enzymes activity implying the critical role of antioxidant in cellular thermotolerance. This clearly demonstrated that TIR can be effectively used for screening high temperature tolerance genotypes in soybean. This study also established the fact that the alternations in antioxidants during induction are vital for imparting tolerance to high temperature stress.