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

Plant Molecular Biology
Hui ZhouYuepeng Han

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 R...Continue Reading

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Citations

Apr 4, 2021·International Journal of Molecular Sciences·Huiling YanXiyang Zhao
Dec 24, 2021·Frontiers in Plant Science·Ya-Ling ZhangZhi-Zhen Fang

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Methods Mentioned

BETA
PCR
two-hybrid
Y2H
Assay
RNA-Seq
transgenic
one-hybrid
electrophoretic

Software Mentioned

MODEL
SWISS

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