The SlWRKY39–SlZF61 module synergistically regulates SlGSTU42 to enhance low‐temperature tolerance in tomato

SUMMARY Low‐temperature stress affects plant growth, and WRKY transcription factors alleviate such damage by regulating downstream genes. This study found that tomato SlWRKY39 significantly responds to low temperatures: its overexpression enhances seedling low‐temperature tolerance by promoting ROS scavenging, while knockout exacerbates ROS accumulation and increases sensitivity to low temperatures. Transcriptome analysis indicated induction of glutathione metabolic pathway genes in slwrky39 plants under low‐temperature stress. Y1H, EMSA, and Dual‐LUC experiments confirmed that SlWRKY39 specifically binds to and activates the SlGSTU42 promoter; silencing SlGSTU42 attenuated the low‐temperature tolerance conferred by SlWRKY39 overexpression, verifying that SlWRKY39 improves low‐temperature tolerance via direct regulation of SlGSTU42 . Additionally, SlZF61 interacts with SlWRKY39, enhancing its regulatory effect on SlGSTU42 . SlZF61 overexpression strengthens low‐temperature tolerance, while knockout increases sensitivity to low temperatures. In summary, under low‐temperature stress, SlWRKY39 and SlZF61 are upregulated expression in tomato; SlWRKY39 binds to the SlGSTU42 promoter, and SlZF61 interacts with SlWRKY39 to form a protein complex, enhancing this binding. They synergistically activate SlGSTU42 transcription, thereby improving seedling low‐temperature tolerance by scavenging ROS. This coordinated regulatory mechanism provides a new theoretical basis and practical insights for enhancing tomato low‐temperature tolerance and ensuring stable production under low‐temperature stress conditions.