VvBBX44 and VvMYBA1 form a regulatory feedback loop to balance anthocyanin biosynthesis in grape

Abstract

Anthocyanins are essential for the quality of perennial horticultural crops, such as grapes. In grapes, ELONGATED HYPOCOTYL 5 (HY5) and MYBA1 are two critical transcription factors that regulate anthocyanin biosynthesis. Our previous work has shown that Vitis vinifera B-box protein 44 (VvBBX44) inhibits anthocyanin synthesis and represses VvHY5 expression in grape calli. However, the regulatory mechanism underlying this regulation was unclear. In this study, we found that loss of VvBBX44 function resulted in increased anthocyanin accumulation in grapevine callus. VvBBX44 directly represses VvMYBA1, which activates VvBBX44. VvMYBA1, but not VvBBX44, directly modulates the expression of grape UDP flavonoid 3- O-glucosyltransferase (VvUFGT). We demonstrated that VvBBX44 represses the transcriptional activation of VvUFGT and VvBBX44 induced by VvMYBA1. However, VvBBX44 and VvMYBA1 did not physically interact in yeast. The application of exogenous anthocyanin stimulated VvBBX44 expression in grapevine suspension cells and tobacco leaves. These findings suggest that VvBBX44 and VvMYBA1 form a transcriptional feedback loop to prevent overaccumulation of anthocyanin and reduce metabolic costs. Our work sheds light on the complex regulatory network that controls anthocyanin biosynthesis in grapevine.

Acknowledgements

We thank professor Yu-Jin Hao (College of Horticulture Science and Engineering, Shandong Agricultural University) for providing the plasmid for the EMSA experiment. All data generated and analyzed in this study are shown in the article or attached as supplementary data. All the materials used in the study are available upon reasonable request from the corresponding author. This work was supported by the National Natural Science Foundation of China (Grant no. U21A20227),and the Strategic Priority Research Program of the Chinese Academy of Sciences (Grant no. XDA23080602). Research conducted as part of the LIA INNOGRAPE International Associated Laboratory.

Author contributions

W.L., H.M., Y.L., S.L., and Y.L. performed the experiments. W.L. and L.W. analyzed the data and wrote the paper. C.R. established the transgenic system of Vitis ‘41B’ cell suspension. W.L. and S.L. designed the research and supervised this study. W.D., P.F., Z.D., Z.L. and L.Y. helped in designing the experiments. Y.Z. and L.Y. provided help in writing. L.W. obtained funding and revised the final version. All authors read and approved the manuscript.

Data availability

The raw RNA-Seq data are archived under accession number GSE98923 in the National Center for Biotechnology Information (NCBI) Sequence Read Archive (SRA). The sequence data can be downloaded from the NCBI, the Genome Database for Rosaceae (GDR) website (https://www.rosaceae.org/NCBI_annotation), and The Arabidopsis Information Resource (TAIR) (https://www.arabidopsis.org/download/index-auto.jsp?dir=%2Fdownload_files%2FProteins%2FAraport11_protein_lists). Additional data supporting the findings of this study are available in the Supplementary Data section of Horticulture Research online article.

Conflict of interest

The authors declare no conflict of interest.

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