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Malate, the predominant organic acid in many fruits, is a crucial component of the organoleptic quality of fruit, including taste and flavor. The genetic and environmental mechanisms affecting malate metabolism in fruit cells have been studied extensively. However, the transcriptional regulation of malate-metabolizing enzymes and vacuolar transporters remains poorly understood. Our previous studies demonstrated that Md MYB1 modulates anthocyanin accumulation and vacuolar acidification by directly activating vacuolar transporters, including Md VHA-B1, Md VHA-E, Md VHP1 and Mdt DT. Interestingly, we isolated and identified a MYB transcription factor, Md MYB73, a distant relative of Md MYB1 in this study. It was subsequently found that Md MYB73 protein bound directly to the promoters of Md ALMT9 (aluminum-activated malate transporter 9), Md VHA-A (vacuolar ATPase subunit A) and Md VHP1 (vacuolar pyrophosphatase 1), transcriptionally activating their expression and thereby enhancing their activities. Analyses of transgenic apple calli demonstrated that Md MYB73 influenced malate accumulation and vacuolar pH. Furthermore, Md CIb HLH1 interacted with Md MYB73 and enhanced its activity upon downstream target genes. These findings help to elucidate how Md MYB73 directly modulates the vacuolar transport system to affect malate accumulation and vacuolar pH in apple. [ABSTRACT FROM AUTHOR]