A Chlorophyll a/b-Binding Protein SlCAB3 Positively Regulates Heat Tolerance Through Interaction with SlDREBA4 in Tomato

High-temperature stress severely limits the growth, development, and productivity of tomatoes. Understanding the molecular mechanisms underlying its thermotolerance is crucial for breeding heat-resistant varieties. This study employed a stepwise experimental strategy to systematically elucidate the role of the chlorophyll a/b-binding protein SlCAB3 in tomato thermotolerance. First, a high-temperature responsive transcription factor, SlDREBA4, previously identified in our lab, was used in a yeast two-hybrid screen to identify potential interacting proteins, including SlCAB3. The interaction between SlDREBA4 and SlCAB3 was further validated using tobacco in vivo luciferase complementation imaging (LCI) and in vitro pull-down assays. Subsequently, the expression patterns of SlCAB3 under heat stress were analyzed, and its biological function was further evaluated through overexpression, gene silencing, and knockout experiments. Additionally, reactive oxygen species (ROS) accumulation, antioxidant enzyme activities, chlorophyll content, and the expression of stress-responsive genes were measured to comprehensively assess their physiological and molecular regulatory roles. The results indicate that SlCAB3 encodes a typical chlorophyll a/b-binding protein and is rapidly induced by heat stress. Overexpression of SlCAB3 significantly enhances plant thermotolerance, evidenced by reduced heat damage, increased chlorophyll content, decreased ROS accumulation, elevated antioxidant enzyme activities, and upregulation of antioxidant-related genes. Conversely, silencing SlCAB3 produces opposite effects. Moreover, co-expression of SlCAB3 with SlDREBA4 further improves thermotolerance, accompanied by enhanced expression of heat shock protein-related and antioxidant-related genes. In conclusion, SlCAB3 is a positive regulator of tomato thermotolerance, and the interaction module formed with SlDREBA4 may collectively enhance heat resistance by strengthening antioxidant defense and heat stress response mechanisms.