| Summary: | The AT-hook motif nuclear-localized (AHL) family members play key roles in plant biological processes via protein–protein and protein-DNA interactions. Here, 22 non-redundant <i>PpAHL</i> genes were identified and analyzed in peach (<i>Prunus persica</i>), one of economically important non-timber forestry crops. The maximum-likelihood (ML) tree classified the <i>PpAHLs</i> into two clades (Clade-A and Clade-B) with three subfamilies: Type_I, Type_II, and Type_III. Exon–intron analysis exhibited that the <i>PpAHLs</i> from Type_I except one (<i>Prupe.1G530300.1</i>) lacked introns, and the <i>PpAHLs</i> from Type_II and Type_III gradually emerged with intron additions, indicating spatial expression patterns, evolutionarily distinct temporal patterns and, likely, neofunctionalization. Duplication event analysis suggested that <i>PpAHLs</i> in peach were mainly expanded through the large-scale duplication events. RNA-seq data showed that <i>PpAHLs</i> were induced by drought stress, and two genes (<i>Prupe.1G530300.1</i> and <i>Prupe.1G034400.1</i>) from Type_I AHLs were induced at all time points, indicating that they might play key roles in the response to drought stress in peach. The tissue-specific expression pattern of <i>PpAHLs</i> exhibited their biological functions in the development of these specific tissues. In addition, the transient overexpression of <i>Prupe.1G530300.1</i> and <i>Prupe.1G034400.1</i> resulted in significant changes in sugar content, suggesting that they may be positive regulators of sugar accumulation in peach fruits. Our study provided novel insights into the roles of <i>PpAHLs</i> in plant development, which was helpful for the functional analysis of peach and related woody fruit trees, and for formulating new strategies for further breeding.
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