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Wheat is the main source of carbohydrates and amino acids consumed in the world. Amino acids and other physiological characters were determined in six wheat genotypes viz. IBWSN-1010, IBWSN -1025, TD-1, ESW-9525, Khirman and Chakwal-86 for pot experiment, to evaluate the response of genotypes under water stress at Nuclear Institute of Agriculture (NIA), Tandojam, Sindh, Pakistan, during 2018. Eight different physiological indices (Proline content, glycine-betaine, total sugars, total chlorophyll, nitrate reductase activity (NRA), potassium (K+) content, osmotic potential (OP) and relative water content (RWC)) were determined. The variance of analysis shows two-way interaction one with water stress [Control (normal four irrigations) and the other with terminal drought (Soaking dose) having significance at p ≤ 0.05. It was observed that, ESW9525, IBWSN-1010 and IBWSN-1025 exhibited the tolerance followed by, Khirman and Chakwal-86 by maintaining their OP and accumulation of higher proline and glycine-betaine content. Whereas, moderate total soluble sugars were found in these genotypes. However, NRA increased in IBWSN-1010 enhancing tolerance under water stress. [ABSTRACT FROM AUTHOR]