Stress adaptation mechanisms involve a series of interconnected responses aimed at maintaining cellular homeostasis. Methylglyoxal (MG), a highly reactive metabolite, is known to increase in response to stress in plants. It is primarily produced as a byproduct of glycolytic and calvin cycle reactions. Owing to its highly reactive nature, MG can readily modify proteins and nucleic acids thereby, altering cellular processes and causing oxidative stress. Despite being cytotoxic, MG can also function as a stress signal molecule, emphasizing the need for maintaining a delicate balance in its levels. Our group has been working towards developing strategies to evaluate and counter the toxic effects of MG for generation of stress resilience in plants. Glyoxalase pathway, consisting of GLYI and GLYII enzymes, serves as the primary route for MG metabolism. By restricting a rise in MG levels through overexpression of glyoxalase pathway, we have shown that adverse effects of MG can be overcome. Interestingly, lowering MG levels in plants provides tolerance to both abiotic and biotic stresses. Additionally, our group has also discovered the presence of GLYIII enzymes in rice which can metabolize MG in a single step. Investigations into the role of GLYIII enzymes unveil their multifaceted functions beyond MG detoxification. Interestingly, glyoxalase I/II and III exist as multi-membered family in plants. Biochemical and molecular studies on multiple members encoding GLYI and GLYII enzymes have unravelled their diverse properties, with some being stress-inducible while others exhibiting constitutive expression and distinct subcellular localization. Furthermore, certain members lack MG detoxification ability but have acquired alternative functions, presenting examples of functional divergence. The tight cellular control of MG levels, evidenced by the presence of multiple pathways for its removal, suggests that plants which restrict a rise in MG levels under stress offer a promising solution for generating climate-ready plants with significantly enhanced stress tolerance.