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Two amino acid changes in the R3 repeat cause functional divergence of two clustered MYB10 genes in peach

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Abstract

R2R3-MYB genes play a pivotal role in regulating anthocyanin accumulation. Here, we report two tandemly duplicated R2R3-MYB genes in peach, PpMYB10.1 and PpMYB10.2, with the latter showing lower ability to induce anthocyanin accumulation than the former. Site-directed mutation assay revealed two amino acid changes in the R3 repeat, Arg/Lys66 and Gly/Arg93, responsible for functional divergence between these two PpMYB10 genes. Anthocyanin-promoting activity of PpMYB10.2 was significantly increased by a single amino acid replacement of Arg93 with Gly93. However, either the Gly93 → Arg93 or Arg66 → Lys66 substitutions alone showed little impact on anthocyanin-promoting activity of PpMYB10.1, but simultaneous substitutions caused a significant decrease. Reciprocal substitution of Arg/Gly93 could significantly alter binding affinity to PpbHLH3, while the Arg66 → Lys66 substitution is predicted to affect the folding of the MYB DNA-binding domain, instead of PpbHLH3-binding affinity. Overall, the change of anthocyanin-promoting activity was accompanied with that of bHLH-binding affinity, suggesting that DNA-binding affinity of R2R3-MYBs depends on their bHLH partners. Our study is helpful for understanding of functional evolution of R2R3-MYBs and their interaction with DNA targets.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (Grant No. 31672134), the China Agriculture Research System (Grant No. CARS-30), and the Overseas Construction Plan for Science and Education Base, China-Africa Center for Research and Education, Chinese Academy of Sciences (SAJC201327).

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Authors and Affiliations

  1. Key Laboratory of Plant Germplasm Enhancement and Specialty Agriculture, Wuhan Botanical Garden of the Chinese Academy of Sciences, Wuhan, 430074, China

    Hui Zhou, Liao Liao, Shengli Xu, Collins Ogutu, Lu Wang & Yuepeng Han

  2. University of Chinese Academy of Sciences, 19A Yuquanlu, Beijing, 100049, China

    Shengli Xu & Collins Ogutu

  3. Institute of Forestry and Pomology, Beijing Academy of Agriculture and Forestry Sciences, Beijing, 100097, China

    Fei Ren, Jianbo Zhao & Quan Jiang

  4. Key Laboratory of Genetic Improvement and Ecophysiology of Horticultural Crops, Horticultural Institute, Anhui Academy of Agricultural Sciences, Hefei, 230031, China

    Hui Zhou

  5. Sino-African Joint Research Center, Chinese Academy of Sciences, Wuhan, 430074, China

    Yuepeng Han

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  1. Hui Zhou
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  2. Liao Liao
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YH and HZ conceived and designed the experiments. HZ, LL, SX, FR, JZ, and LW performed the experiments. YH and HZ wrote the paper. CO and QJ revised the manuscript.

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Correspondence to Yuepeng Han.

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Zhou, H., Liao, L., Xu, S. et al. Two amino acid changes in the R3 repeat cause functional divergence of two clustered MYB10 genes in peach. Plant Mol Biol 98, 169–183 (2018). https://doi.org/10.1007/s11103-018-0773-2

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