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Heat stress due to increased temperature is an agricultural problem in many areas in the world. It affects the wide spectrum of both biochemical and physiological responses within the plant cells. Relative Leaf Water Content (RLWC) and Water Saturation Deficit (WSD) was evaluated under control (25±2°C), heat stress (35±2°C and 40±2°C) and interactive effect of heat stress and trehalose was studied in wheat (Triticum aestivum L.) genotypes viz. HD2967, C306, PBW621, PBW343, PBW175 and PBW590. Trehalose at concentration of 1mM and 1.5mM was applied followed by heat stress of 35±2°C (moderate) and 40±2°C (severe) for 4 and 8 hours. Heat stress reduced the RLWC and increased the WSD of studied wheat genotypes. The duration of heat stress for 8 hours had more adverse effect on tested parameters. Trehalose application ameliorated the adverse effect of heat stress to some extent. Trehalose application @1.5mM concentration was found more effective as compared with 1mM concentration tested presently.