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Differential anthocyanin accumulation in radish taproot: importance of RsMYB1 gene structure

  • Biao Lai
  • Yuanyi Cheng
  • Hong Liu
  • Qiuxia Wang
  • Qi Wang
  • Chunlan Wang
  • Rui Su
  • Fabo Chen
  • Huicong WangEmail author
  • Lina DuEmail author
Original Article

Abstract

Key message

RsMYB1a was the crucial MYB, and RsbHLH4 is the essential partner in regulating the anthocyanin biosynthesis in radish.

Abstract

There are four color types of radish according to whether or not the anthocyanin accumulates in the skin and flesh of taproot. Red radishes accumulate a substantial amount of anthocyanins in both the skin and flesh. It is well known that the MYB-bHLH-WD40 transcription factor(s) complex regulates the biosynthesis of anthocyanin in plants. Here in, four candidate MYB and bHLH genes, RsMYB1a, RsMYB1b, RsbHLH2 and RsbHLH4, were isolated from red radish ‘Hongxin 1’. The expression of RsbHLH4 and the two structural genes RsANS and RsUFGT was significantly positively correlated with anthocyanin contents. The expression of RsMYB1a was also highly correlated with anthocyanin accumulation, particularly when the white flesh sample of ‘Hongxin 1-1’ was excluded. The transient expression of RsMYB1a in the radish cotyledon and leaf induced anthocyanin accumulation with even stronger promoting role when expression in combination with RsbHLH4. These results suggested that RsMYB1a was the crucial MYB, and that RsbHLH4 is an essential partner in regulating the biosynthesis of anthocyanins in radish. The low or undetectable RsbHLH4 expression paralleled the lack of anthocyanin accumulation in the white flesh of ‘Hongxin 1-1’ and ‘Shaguan 1’. Assays demonstrated that RsMYB1a interacted with RsbHLH4 and activated the expression of RsbHLH4. Notably, all the dark red radish cultivars have a longer RsMYB1a genomic DNA sequence, while the short and nonfunctional RsMYB1a is present in non-red cultivars. The length of the first intron and the presence of an early stop codon of RsMYB1 might underlie the differential anthocyanin accumulation in the radish taproot.

Keywords

Red radish Anthocyanins MYB bHLH Transcriptional regulation 

Abbreviations

4CL

4-Coumarate CoA ligase

AbA

Aureobasidin A

ANS

Anthocyanidin synthase

BiFC

Bimolecular fluorescence complementation

C4H

Cinnamate 4-hydroxylase

CHI

Chalcone isomerase

CHS

Chalcone synthase

DFR

Dihydroflavonol reductase

F3H

Flavanone 3-hydroxylase

ORF

Open read frame

PAL

Phenylalanine ammonia lyase

qRT-PCR

Quantitative real-time PCR

SNPs

Single-nucleotide polymorphisms

UFGT

UDP-glucose:flavonoid 3-O-glucosyltransferase

YFP

Yellow fluorescence protein

Notes

Acknowledgements

This study was supported in part by Scientific and Technological Research Program of Chongqing Municipal Education Commission (Grant nos. KJ1712303 and KJ1712302), Chongqing Natural Science Foundation (Grant nos. cstc2017jcyjA0001, cstc2016jcyjA0136 and cstc2017shms-xdny80074) and Yangtze Normal University (Grant nos. 2016KYQD20 and 2016XJQN06).

Author contribution statement

BL and LND designed the study. BL, YYC, HL, QW, CLW, and RS carried out the experiments. BL and LND analyzed the data. FBC sampled the red radishes. BL, LND and HCW wrote the manuscript. All the authors read and agreed to submit the manuscript.

Compliance with ethical standards

Conflict of interest

All the authors declare no conflict of interest.

Supplementary material

299_2019_2485_MOESM1_ESM.docx (3.7 mb)
Supplementary material 1 (DOCX 3820 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Biao Lai
    • 1
  • Yuanyi Cheng
    • 1
  • Hong Liu
    • 1
  • Qiuxia Wang
    • 1
  • Qi Wang
    • 1
  • Chunlan Wang
    • 1
  • Rui Su
    • 1
  • Fabo Chen
    • 1
  • Huicong Wang
    • 1
    • 2
    Email author
  • Lina Du
    • 1
    Email author
  1. 1.School of Advanced Agriculture and BioengineeringYangtze Normal UniversityChongqingPeople’s Republic of China
  2. 2.College of HorticultureSouth China Agricultural UniversityGuangzhouPeople’s Republic of China

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