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Differential activation of anthocyanin biosynthesis in Arabidopsis and tobacco over-expressing an R2R3 MYB from Chinese bayberry

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Abstract

MrMYB1, an R2R3 MYB transcription factor (TF) gene associated with anthocyanin biosynthesis in Chinese bayberry (Myrica rubra Sieb. and Zucc.), was introduced into Arabidopsis and tobacco (Nicotiana tabacum) under the control of the CaMV 35S promoter. Overexpression of MrMYB1 induced anthocyanin accumulation in all tissues of Arabidopsis as well as in petals, ovaries and young seeds of tobacco, but not in tobacco leaves. The anthocyanin biosynthetic pathway, including chalcone synthase (CHS), dihydroflavonol 4-reductase (DFR) and anthocyanidin synthase (ANS), and the basic helix-loop-helix (bHLH) transcriptional partner TRANSPARENT TESTA8 (TT8), were up-regulated significantly in MrMYB1-overexpressing Arabidopsis. In MrMYB1-overexpressing tobacco petals, ovaries and young seeds, anthocyanin biosynthetic genes and bHLH partners NtAn1a and NtAn1b, were up-regulated. In contrast, high expression of MrMYB1 in transgenic tobacco leaves did not induce the expression of anthocyanin biosynthesis. Unlike in petals and ovaries, the foliar transcript level of NtAn1a and NtAn1b was extremely low and not stimulated by MrMYB1 transformation. These results show that higher expression of an endogenous bHLH partner, either intrinsically or stimulated by exogenous gene transformation, is required for anthocyanin production in plant tissues, and the different abundance in endogenous bHLH transcript accounts for differential accumulation of anthocyanin in Arabidopsis and tobacco leaves. These findings demonstrate that higher levels of expression of an endogenous bHLH partner, either intrinsically or following genetic transformation, are required for anthocyanin production in plant tissues. Moreover, differences in levels of endogenous bHLH transcripts account for observed differential accumulation of anthocyanin in leaves of Arabidopsis and tobacco.

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Abbreviations

TF:

Transcription factor

CaMV:

Cauliflower mosaic virus

CHS:

Chalcone synthase

DFR:

Dihydroflavonol 4-reductase

ANS:

Anthocyanidin synthase

bHLH:

Basic helix-loop-helix

TT8:

TRANSPARENT TESTA8

PAP1:

Production of Anthocyanin Pigment 1

ANT1:

Anthocyanin1

CDS:

Coding sequence

RNR2:

Ribonucleotide reductase R2

NPT II:

Neomycin phosphotransferase II

HPLC:

High-performance liquid chromatography

CTAB:

Cetyltrimethylammonium bromide

qPCR:

Real-time quantitative PCR

CHI:

Chalcone isomerase

UFGT:

UDP-glucose: flavonoid 3-O-glucosyltransferase

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Acknowledgments

This research was supported by the National High Technology Research and Development Program of China [2013AA102606], the National Natural Science Foundation of China [31071781], the Program of International Science and Technology Cooperation [2011DFB31580], the Special Scientific Research Fund of Agricultural Public Welfare Profession of China (201203089), the Science and Technology Project of Zhejiang Province (2009C14023), and the Specialized Research Fund for the Doctoral Program of Higher Education of China (20090101110099).

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Authors and Affiliations

  1. Laboratory of Fruit Quality Biology/The State Agriculture Ministry Laboratory of Horticultural Plant Growth, Development and Quality Improvement, Zhejiang University, Hangzhou, 310058, People’s Republic of China

    Yu-Ji Huang, Song Song, Xiao-Fen Liu, Xue-Ren Yin, Chang-Jie Xu & Kun-Song Chen

  2. The New Zealand Institute of Plant and Food Research, Private Bag 92169, Auckland, New Zealand

    Andrew C. Allan

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  1. Yu-Ji Huang
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  2. Song Song
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  4. Xiao-Fen Liu
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Correspondence to Chang-Jie Xu.

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Huang, YJ., Song, S., Allan, A.C. et al. Differential activation of anthocyanin biosynthesis in Arabidopsis and tobacco over-expressing an R2R3 MYB from Chinese bayberry. Plant Cell Tiss Organ Cult 113, 491–499 (2013). https://doi.org/10.1007/s11240-013-0291-5

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