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A previous study showed that foliar application of ethephon to litchi trees with mature shoots and dormant terminal buds during autumn successfully inhibited new vegetative shoot growth prior to floral induction thereby promoting carbohydrate accumulation and flowering. However, the functional mechanisms of ethylene, the breakdown product of ethephon, in the leaves and terminal buds of litchi and its involvement in the flowering process is largely unknown. Therefore, this study aimed to investigate the phenological, physiological and molecular changes underlying ethephon application and its associations with bud dormancy and flowering in litchi. Ethephon was applied as a single full canopy spray at a concentration of 1000 mg⋅L−1 to 'Mauritius' litchi trees with mature vegetative shoots and dormant terminal buds during late autumn of 2018 (mid-April; Southern Hemisphere). Untreated trees served as a control. Phenological characteristics, such as bud dormancy and panicle development, leaf chlorophyll (as an indicator of shoot maturity), ethylene evolution, gene expression levels of flowering- (LcFT2 , LcFLC and LcAP1), dormancy- (LcSVP1 and LcSVP2) and ethylene pathway-related (LcEIN3) genes and non-structural carbohydrates were determined in terminal buds, leaves and/or shoots. Ethephon application induced bud dormancy, significantly delayed panicle emergence and promoted pure floral panicle development under more favorable inductive conditions. Ethylene evolution increased sharply 2 h after application in both leaves and terminal buds, but decreased rapidly thereafter in the leaves, while remaining high in terminal buds for seven days before gradually declining. Ethephon application significantly increased relative expression of LcEIN3 and LcFLC in terminal buds one day after application, while LcFT2 expression in leaves and LcAP1 expression in terminal buds were significantly increased at the bud break stage. Significant treatments differences were also observed for various carbohydrate metabolites in leaves and shoots at the bud break or floral initiation stage. Our study provided evidence that ethephon application plays an important role in the physiological and molecular regulation of bud dormancy of litchi. By influencing the time of bud break, ethephon application can be a useful tool to manage panicle emergence under less inductive conditions. [ABSTRACT FROM AUTHOR]