| Summary: | Sugars and organic acids significantly impact fruit sensory quality, but their accumulation patterns and regulatory mechanisms during the development of <i>Rosa roxburghii</i> fruit are still unclear. We utilized transcriptomics and metabolomics to investigate genes related to sugar and organic acid metabolism in <i>Rosa roxburghii</i>. Metabolomics data revealed that sucrose, glucose and fructose were the primary sugars, whereas citric acid and malic acid were the primary organic acids in <i>Rosa roxburghii</i> fruit. We constructed the metabolic pathways of major sugars and organic acids in <i>Rosa roxburghii</i> and identified five key genes involved in sugar and organic acid synthesis. In addition, we identified a module containing 132 transcription factors that was significantly associated with sucrose, citric acid and malic acid. Based on quantitative polymerase chain reaction (qPCR), we identified 13 transcription factors involved in sugar and organic acid metabolism, including the transcription factor <i>RrANL2</i> and the sucrose synthase gene <i>RrSUS3</i>. Further yeast one-hybrid and dual luciferase assays showed that <i>RrANL2</i> could bind to the promoter of <i>RrSUS3</i> to increase its expression. These results provide new insights into the metabolism of sugars and organic acids in <i>Rosa roxburghii</i> fruit.
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