Exogenous 5-aminolevulinic acid promotes carotenoid accumulation in tomato fruits by regulating ethylene biosynthesis and signaling

5-Aminolevulinic acid (ALA) can not only improve fruit yield and quality, but also increase the lycopene content in tomato fruits. Furthermore, ALA has been shown to promote system-2 ethylene production in tomato fruits. However, the specific interactions between ALA and ethylene during fruit ripening remain unclear. In this study, we treated tomato fruits with ALA, 1-aminocyclopropane-1-carboxylic acid (ACC), aminooxyacetic acid (AOA) + AgNO3, and AOA + AgNO3 + ALA and analyzed ethylene emissions, carotenoid contents, and the relative gene expression levels related to fruit ripening, carotenoid contents, ethylene synthesis, and signal transduction. The ALA treatment significantly enhanced ethylene bursts and carotenoid accumulation, and significantly upregulated the expression of ethylene and carotenoid-related genes, such as SlACS2, SlACS4, SlACO1, SlPSY1, and SlPDS. We also observed that the gene expression levels associated with carotenoid synthesis were downregulated in fruits treated with a combination of ethylene inhibitors (AOA + AgNO3). However, there was a significant upregulation in the gene expression levels associated with carotenoid synthesis and an increase in carotenoid content when fruits were treated with AOA + AgNO3 + ALA. After silencing SlACO1 expression, the total carotenoid content and SlPSY1 expression decreased significantly, while this effect was reversed after exogenous application of ALA. These results indicated that ALA promotes carotenoid accumulation in tomato fruits by promoting ethylene biosynthesis. In conclusion, our results highlighted the role of ALA in promoting carotenoid accumulation and ripening in tomato fruits by regulating ethylene synthesis, thereby providing a novel strategy for improving fruit quality.