CRISPR-CAS9–MEDIATED DEVELOPMENT OF CLIMATE-RESILIENT WHEAT GENOTYPES UNDER HEAT AND DROUGHT STRESS CONDITIONS IN PAKISTAN

Abstract

Wheat production in Pakistan is increasingly threatened by heat and drought stress induced by climate change, leading to significant yield instability and food security concerns. This study investigated the potential of CRISPR-Cas9–mediated genome editing for developing climate-resilient wheat genotypes with enhanced tolerance to abiotic stress conditions. A quantitative experimental design was employed under controlled greenhouse conditions, where CRISPR-edited wheat lines (single-gene and multi-gene) were evaluated against non-edited control genotypes under simulated heat and drought stress environments. Physiological, biochemical, and agronomic traits including photosynthetic rate, chlorophyll content, water-use efficiency, biomass accumulation, and grain yield were analyzed using ANOVA, correlation, and regression techniques. The results revealed that CRISPR-edited wheat genotypes significantly outperformed control plants across all measured traits, with multi-gene edited lines exhibiting the highest stress tolerance and yield stability. Statistical analysis confirmed significant genotype and stress interaction effects, indicating enhanced adaptability of edited lines under adverse conditions. The findings demonstrate that CRISPR-Cas9 technology substantially improves wheat resilience by enhancing physiological efficiency and stabilizing yield under extreme environmental stress. The study concludes that genome editing represents a promising strategy for sustainable wheat production in climate-vulnerable regions such as Pakistan.