Anthocyanin-enriched Chinese cabbage is popular due to its attractive color and health-enhancing antioxidants. Anthocyanin accumulation is controlled by several regulators, but little information is available on hierarchical interactions between activators and repressors. Here, we characterized an R2R3-MYB repressor in Chinese cabbage (Brassica rapa), designated BrMYBR1, which negatively regulates anthocyanin biosynthesis. BrMYBR1 is highly expressed under sucrose supplementation and high-light intensity conditions that can induce anthocyanin accumulation. Transgenic tobacco (Nicotiana tabacum) heterologously expressing BrMYBR1 showed reductions in anthocyanin and transcripts of anthocyanin biosynthesis genes in flowers, revealing that BrMYBR1 represses anthocyanin biosynthesis. Nucleus-localized BrMYBR1 interacts with a basic helix-loop-helix protein, TRANSPARENT TESTA 8 (BrTT8), but not with the R2R3-MYB protein PRODUCTION OF ANTHOCYANIN PIGMENT 1 (BrPAP1). Additionally, transient infiltration assays in Chinses cabbage cotyledons and tobacco leaves revealed that BrMYBR1 represses pigment accumulation by inhibiting anthocyanin biosynthesis gene transcription through its C-terminal C1 and C2 motifs. Transient promoter activation assays in Chinese cabbage cotyledon protoplasts demonstrated that BrMYBR1 represses the transactivation of CHALCONE SYNTHASE and DIHYDROFLAVONOL REDUCTASE by BrPAP1 and BrTT8. Moreover, BrMYBR1 was induced upon co-expression with BrPAP1 and BrTT8, and BrMYBR1 then performed feedback regulation of BrPAP1 and BrTT8 expression. Together, these results demonstrated that BrMYBR1 hindered the formation of an active MYB–bHLH–WD40 (MBW) complex, thereby inhibiting anthocyanin biosynthesis, in addition to directly repressing the promoters of anthocyanin biosynthesis genes. Our findings present a model whereby anthocyanin biosynthesis is balanced by the hierarchical regulation between activator and repressor and clarify how anthocyanin biosynthesis is fine-tuned in Chinese cabbage.