miR156b-targeted VvSBP8/13 functions downstream of the abscisic acid signal to regulate anthocyanins biosynthesis in grapevine fruit under drought

Abstract

Anthocyanins are the primary color components of grapevine berries and wines. In cultivation practices, a moderate water deficit can promote anthocyanin accumulation in red grape skins. Our previous study showed that abscisic acid (ABA) plays a key role in this process. Herein, we identified a microRNA, vv-miR156b, that is generated in grapevine berries in response to drought stress, along with increasing anthocyanin content and biosynthetic structural gene transcripts. In contrast, vv-miR156b short tandem target mimic (STTM) function-loss callus exhibits the opposite phenotype. Results from in vivo and in vitro experiments revealed that the ABA-signaling-regulated transcription factor VvAREB2 binds directly to the ABA-responsive element (ABRE) of the MIR156b promoter and activates miR156b expression. Furthermore, two miR156b downstream targets, VvSBP8 and VvSBP13, exhibited reduced grape anthocyanin content in their overexpressors but there was a contrary result in their CRISPR-edited lines, the decrease in anthocyanin content was rescued in miR156b and SBP8/13 double overexpressors. We further demonstrated that both VvSBP8 and VvSBP13, encoding transcriptional repressors, displayed sufficient ability to interact with VvMYC1 and VvMYBA1, thereby interfering with MYB-bHLH-WD (MBW) repeat transcriptional complex formation, resulting in the repression of anthocyanin biosynthesis. Our findings demonstrate a direct functional relationship between ABA signaling and the miR156-SBP-MBW complex regulatory module in driving drought-induced anthocyanin accumulation in grape berries.

Acknowledgements

This work was supported by the National Natural Science Foundation of China (grants 32371924, 32260727, and 31801811), the Key Research and Development Program of Shaanxi Province (grant 2023-GHZD-17), the Key Research and Development Programme of Xinjiang Province (grant 2022B02034-3), and the China Agriculture Research System (grant CARS-29-zp-6). We are grateful to Dr Jing Zhang, Dr Yangyang Yuan, Miss Haiying Wang, and Miss Beibei He (Horticulture Science Research Center, Northwest A&F University, Yangling, China) for providing professional technical assistance; We also thank Prof. Yaoguang Liu (South China Agricultural University), Prof. Mingzhi Yang (Yunnan University), Prof. Dongqing Xu (Nanjing Agricultural University), and Prof. Chao Ma (China Agricultural University) for gifting us materials and plasmids.

Author contributions

T.F.X., J.F.M., and Z.W.Z. conceived and designed the experiments. S.H.G., M.Z., M.X.F., G.P.L., X.Q.T., and R.H.R. performed the experiments, and S.H.G. and M.Z. analyzed the data. S.H.G., T.F.X., M.Z., L.T., Y.L.F., and J.F.M. wrote and edited the manuscript. All authors reviewed and approved the manuscript.

Data availability

All relevant data generated or analyzed are included in the manuscript and the supporting materials. The sequence information used in this study can be obtained from the NCBI database (https://www.ncbi.nlm.nih.gov/). The GenBank accession numbers are as follows: VvAREB2 (XM_010655778.2), VvUFGT (XM_002276999.4), VvLDOX (NM_001281218.1), VvDFR (XM_002281822), VvSBP8 (XM_002278476.1), VvSBP13 (XM_002274466.1), VvACTIN (XM_002282480.4), VvUBI (XM_002273532.2), and AtACTIN (AT3G18780).

Conflict of interest

None declared.

Supplementary data

Supplementary data is available at Horticulture Research online.

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