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Characterization of an apple TT2-type R2R3 MYB transcription factor functionally similar to the poplar proanthocyanidin regulator PtMYB134

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Abstract

Main conclusion

The apple MdMYB9 gene encodes a positive regulator of proanthocyanidin synthesis that activates anthocyanidin reductase promoters from apple and poplar via interaction with basic helix-loop-helix proteins.

The regulation of proanthocyanidins (PAs, condensed tannins) is of great importance in food plants due to the many benefits of PAs in the human diet. Two candidate flavonoid MYB regulators, MdMYB9 and MdMYB11, were cloned from apple (Malus × domestica) based on their similarity to known MYB PA regulators. Transcript accumulation of both MdMYB9 and MdMYB11 was induced by high light and wounding, similar to the poplar (Populus spp) PA regulator PtMYB134. In transient activation assays with various basic helix-loop-helix (bHLH) co-regulators, MdMYB9 activated apple and poplar anthocyanidin reductase (ANR) promoters, while MdMYB11 showed no activity. Potential transcription factor binding elements were found within several ANR promoters, and the importance of the bHLH binding site (E-box) on ANR promoter activation was demonstrated via mutational analysis. The ability of MdMYB9 and PtMYB134 to reciprocally activate ANR promoters from both apple and poplar and to partner with heterologous bHLH co-factors from these plants confirms the high degree of conservation of PA regulatory complexes across species. The similarity in apple and poplar PA regulation suggests that regulatory genes from poplar could be effectively employed for metabolic engineering of the PA pathway in apple.

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Abbreviations

ANR:

Anthocyanidin reductase

PA:

Proanthocyanidin

bHLH:

Basic helix-loop-helix

TT2:

TRANSPARENT TESTA 2

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Acknowledgments

We thank Dr. Roger Hellens for the gift of the pGreen Luc800 vectors, and Dr. Maasaki Sakuta for the AtBAN promoter. We also acknowledge Okanagan Specialty Fruits Inc. for providing plant material and support, Brad Binges for help in maintaining plants, and Tieling Zhang for technical assistance. This work was funded by the Natural Sciences and Engineering Research Council of Canada (NSERC) in the form of a Strategic Project Grant.

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Correspondence to C. Peter Constabel.

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Gesell, A., Yoshida, K., Tran, L.T. et al. Characterization of an apple TT2-type R2R3 MYB transcription factor functionally similar to the poplar proanthocyanidin regulator PtMYB134. Planta 240, 497–511 (2014). https://doi.org/10.1007/s00425-014-2098-y

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Keywords

  • Gene expression
  • Transcription factor
  • Promoter
  • Flavonoids
  • Condensed tannins