Plant Cell Reports

, Volume 35, Issue 3, pp 641–653 | Cite as

Activation of anthocyanin biosynthesis by expression of the radish R2R3-MYB transcription factor gene RsMYB1

  • Sun-Hyung LimEmail author
  • Ji-Hye Song
  • Da-Hye Kim
  • Jae Kwang Kim
  • Jong-Yeol Lee
  • Young-Mi Kim
  • Sun-Hwa HaEmail author
Original Article


Key message

RsMYB1, a MYB TF of red radish origin, was characterized as a positive regulator to transcriptionally activate the anthocyanin biosynthetic machinery by itself in Arabidopsis and tobacco plants.


Anthocyanins, providing the bright red-orange to blue-violet colors, are flavonoid-derived pigments with strong antioxidant activity that have benefits for human health. We isolated RsMYB1, which encodes an R2R3-MYB transcription factor (TF), from red radish plants (Raphanus sativus L.) that accumulate high levels of anthocyanins. RsMYB1 shows higher expression in red radish than in common white radish, in both leaves and roots, at different growth stages. Consistent with RsMYB1 function as an anthocyanin-promoting TF, red radishes showed higher expression of all six anthocyanin biosynthetic and two anthocyanin regulatory genes. Transient expression of RsMYB1 in tobacco showed that RsMYB1 is a positive regulator of anthocyanin production with better efficiency than the basic helix-loop-helix (bHLH) TF gene B-Peru. Also, the synergistic effect of RsMYB1 with B-Peru was larger than the effect of the MYB TF gene mPAP1D  with B-peruArabidopsis plants stably expressing RsMYB1 produced red pigmentation throughout the plant, accompanied by up-regulation of the six structural and two regulatory genes for anthocyanin production. This broad transcriptional activation of anthocyanin biosynthetic machinery in Arabidopsis included up-regulation of TRANSPARENT TESTA8, which encodes a bHLH TF. These results suggest that overexpression of RsMYB1 promotes anthocyanin production by triggering the expression of endogenous bHLH genes as potential binding partners for RsMYB1. In addition, RsMYB1-overexpressing Arabidopsis plants had a higher antioxidant capacity than did non-transgenic control plants. Taken together, RsMYB1 is an actively positive regulator for anthocyanins biosynthesis in radish plants and it might be one of the best targets for anthocyanin production by single gene manipulation being applicable in diverse plant species.


Anthocyanins Antioxidant R2R3-MYB Radish 



4-coumarate-CoA ligase


basic helix-loop-helix


anthocyanidin synthase


chalcone isomerase


chalcone synthase


dihydroflavonol 4-reductase


flavanone 3-hydroxylase


flavonoid 3′-hydroxylase


phenylalanine ammonia lyase


quantitative real time polymerase chain reaction


transcription factor









This work was supported by a fund from the National Academy of Agricultural Science (PJ01002701) and a grant from the Next-Generation BioGreen 21 Program (PJ01109402), Rural Development Administration, Republic of Korea.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

299_2015_1909_MOESM1_ESM.tif (1.9 mb)
Supplementary Figure legends, Figure 1. The genomic structure of radish RsMYB1 and multiple alignment of the amino acid sequences among R2R3-MYBs. (A) Schematic representation of the structure of RsMYB1. The black boxes represent exons and solid lines represent introns. (B) Multiple sequence alignment of the R2R3-MYB transcription factors related to anthocyanin production that were used in the neighbor-joining phylogenetic tree (Fig. 1). Arrows indicate specific residues that contribute to a motif implicated in the interaction with the bHLH co-factor in Arabidopsis. The red box and black box represent the conserved ANDV motif and the C-terminal-conserved KPRPR[S/T]F motif of anthocyanin-promoting MYBs, respectively. Numbers on the right denote the number of amino acid residues. Residues identical for all the sequences in a given position are in white text on a black background and residues identical for at least over 80 % in a given position are presented in white text on a dark grey background and residues identical ranged from 60 to 80 % in given position are presented in white text on a light gray background. The following sequences were included in the analysis: AmROSEA1 (ABB83826) and AmVENOSA (ABB83828) in Antirrhinum majus, AtPAP1 (NP_176057) and AtPAP2 (NP_176813) in Arabidopsis, BoMYB2 (ADP76650) in Brassica oleracea, GhMYB10 (CAD87010) in Gerbera hybrida, LeANT1 (AAQ55181) in Lycopersicon esculentum, MdMYB10 (ABB84753) in Malus  ×  domestica, OsC1 (CAA75509) in Oryza sativa, PhAN2 (AAF66727) in Petunia  ×  hybrida, RsMYB1 (KR706195) in Raphanus sativus and ZmPl (AAA19821) and ZmC1 (P10290) in Zea mays (TIFF 1926 kb)


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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Sun-Hyung Lim
    • 1
    Email author
  • Ji-Hye Song
    • 1
  • Da-Hye Kim
    • 1
  • Jae Kwang Kim
    • 2
  • Jong-Yeol Lee
    • 1
  • Young-Mi Kim
    • 1
  • Sun-Hwa Ha
    • 3
    Email author
  1. 1.National Academy of Agricultural Science, Rural Development AdministrationJeonjuRepublic of Korea
  2. 2.Division of Life Sciences and Bio-Resource and Environmental CenterIncheon National UniversityIncheonRepublic of Korea
  3. 3.Department of Genetic Engineering and Graduate School of BiotechnologyKyung Hee UniversityYonginRepublic of Korea

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