Engineering Thermal Stress Resistance in Crops Using Protein Solutions and Synthetic Biology

Climate change poses a major threat to global food security by increasing the frequency and severity of extreme temperatures, which can significantly damage crop productivity.To address this challenge, this study explores a synthetic biologybased approach to engineer thermal stress resistance in crops using protective proteins.Specifically, we investigate the roles of heat shock protein HSP70 and trehalose biosynthesis enzymes OtsA and OtsB in enhancing cellular stability under heat and cold stress.Using E. coli as a model system, we constructed two plasmids: pHot, a heat-inducible system for HSP70 expression, and pCold, a cold-inducible system for OtsA/OtsB expression.These plasmids were introduced into E. coli BL21 STAR (DE3), and protein expression was tested under thermal stress (42C and 20C).Western blotting and fluorescence microscopy confirmed successful expression of the target proteins, although heat-inducible dsRED expression was not detected-likely due to inefficient T2A cleavage.Cold-inducible constructs showed strong EGFP fluorescence, supporting the role of OtsA and OtsB in cold tolerance.This work provides a foundation for developing temperature-responsive genetic tools in agricultural biotechnology and informs future strategies for building climate-resilient crops.