Abstract
Key message
Overexpression of CpbHLH1 in Arabidopsis and tobacco resulted in a dramatic decrease in anthocyanin accumulation by repressing the expression of late biosynthesis genes in the flavonoid biosynthesis pathway.
Abstract
Many basic helix–loop–helix (bHLH) transcription factors (TFs) of subgroup IIIf have been characterized as anthocyanin-associated activators in higher plants, but information regarding bHLH TFs that inhibit anthocyanin accumulation remains scarce. In this study, the subgroup IIIf bHLH TF CpbHLH1 from Chimonanthus praecox (L.) was identified as a negative regulator of anthocyanin accumulation. Our results showed that overexpression of CpbHLH1 in model plant species, Arabidopsis and tobacco, resulted in a dramatic decrease in anthocyanin content, whereas the content of proanthocyanidin was little affected. Quantitative RT-PCR (qRT-PCR) assays of the structural genes in the flavonoid biosynthesis pathway revealed that CpbHLH1 inhibits anthocyanin accumulation mainly through repressing the expression of late biosynthesis genes (LBGs). Interactions between CpbHLH1 protein and AtPAP1/NtAN2 protein were detected via yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays. This is the first bHLH repressor of anthocyanin biosynthesis identified in dicotyledons. These results can help us better understand the anthocyanin regulatory network in plants and may provide insights into the diverse functions of bHLH proteins.
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Acknowledgements
This work was funded by the National Natural Science Foundation of China (Nos: 31272207 and 31972442). We thank Professor Wang-jin Lu (State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, College of Horticulture, South China Agricultural University) and Professor Jin-Long Qiu (State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences) for the generous gifts of the pBD vectors and the pap1-D mutant of Arabidopsis, respectively. We are also grateful to Professor Guogui Ning (Key Laboratory of Horticultural Plant Biology, College of Horticulture and Forestry Sciences, Huazhong Agricultural University) for helpful comments on the manuscript.
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RZ, XS and KZ: designed the experiments; RZ: performed the majority of experiments; XS, NY, and LC: supplied the materials; RZ, XS, and KZ: analyzed experimental results. RZ: prepared the initial manuscript draft; LX, XL, and KZ: produced the final version of the manuscript. All authors approved the final manuscript.
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Communicated by Salim Al-Babili.
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Zhao, R., Song, X., Yang, N. et al. Expression of the subgroup IIIf bHLH transcription factor CpbHLH1 from Chimonanthus praecox (L.) in transgenic model plants inhibits anthocyanin accumulation. Plant Cell Rep 39, 891–907 (2020). https://doi.org/10.1007/s00299-020-02537-9
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DOI: https://doi.org/10.1007/s00299-020-02537-9