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Narrow genetic variation limits the success of crop improvement programs. Mutagenesis using ethyl methanesulphonate (EMS) provides an opportunity to increase genetic variation to enhance selection in wheat improvement. This study aimed at establishing the optimum dosage and treatment conditions for EMS to induce genetic variation for drought tolerance and enhanced carbon sequestration in wheat. Seeds of three genotypes (LM29, LM43 and LM75) were treated with four EMS dosages (0, 0.1, 0.4 and 0.7% v/v) at three temperatures (25, 30 and 35 °C) for three exposure periods (1, 1.5 and 2 h) using three replicates. The ideal treatment conditions for effective mutagenesis were 0.7% EMS for 2 h at 35 °C for genotypes LM29 and LM43 and 0.4% EMS for 2 h at 25 °C for LM75. The estimated EMS dosages for LM43, LM29 and LM75 were 0.32%, 1.07% and 1.81% v/v EMS, respectively. This …