The plant hormone ethylene is indispensable to the ripening of climacteric fruits. Although extensive studies have been conducted on ethylene signaling, the ethylene response factor (ERF)-mediated transcriptional regulation of ethylene biosynthesis in pear fruits remains to be fully elucidated. We here constructed, sequenced, and analyzed transcriptome libraries in ethephon-treated and 1-MCP-treated Cuiguan pear fruits. In total, 721 fruit ripening-associated differentially expressed genes were identified. Among them, two key genes exhibited positive correlations: the 1-aminocyclopropane-1-carboxylic acid synthase (ACS)-encoding gene PbrACS3 and the ERF-encoding gene named PbrERF114. PbrERF114 overexpression increased ethylene production as well as the PbrACS3 expression level. Conversely, virus-induced gene silencing downregulated PbrERF114, thereby decreasing ethylene production and reducing PbrACS3 expression levels. Notably, PbrERF114 could directly interact with PbrACS3 and PbrERF24 promoters, thus inducing their expression. However, it did not result in an enhancement in luciferase activity, which is regulated by the PbrACS1b or PbrACO1 promoter. PbrERF24 could directly bind to PbrACO1 as well as PbrACS3 to promote ethylene synthesis. In conclusion, PbrERF114 can directly and indirectly mediate ethylene biosynthesis by transcriptionally regulating PbrACS3 and PbrERF24, respectively, thereby triggering a signaling cascade that induces the expression of both PbrACS3 and PbrACO1.
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