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Characterization of the regulatory network of BoMYB2 in controlling anthocyanin biosynthesis in purple cauliflower

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Abstract

Purple cauliflower (Brassica oleracea L. var. botrytis) Graffiti represents a unique mutant in conferring ectopic anthocyanin biosynthesis, which is caused by the tissue-specific activation of BoMYB2, an ortholog of Arabidopsis PAP2 or MYB113. To gain a better understanding of the regulatory network of anthocyanin biosynthesis, we investigated the interaction among cauliflower MYB–bHLH–WD40 network proteins and examined the interplay of BoMYB2 with various bHLH transcription factors in planta. Yeast two-hybrid studies revealed that cauliflower BoMYBs along with the other regulators formed the MYB–bHLH–WD40 complexes and BobHLH1 acted as a bridge between BoMYB and BoWD40-1 proteins. Different BoMYBs exhibited different binding activity to BobHLH1. Examination of the BoMYB2 transgenic lines in Arabidopsis bHLH mutant backgrounds demonstrated that TT8, EGL3, and GL3 were all involved in the BoMYB2-mediated anthocyanin biosynthesis. Expression of BoMYB2 in Arabidopsis caused up-regulation of AtTT8 and AtEGL3 as well as a subset of anthocyanin structural genes encoding flavonoid 3′-hydroxylase, dihydroflavonol 4-reductase, and leucoanthocyanidin dioxygenase. Taken together, our results show that MYB–bHLH–WD40 network transcription factors regulated the bHLH gene expression, which may represent a critical feature in the control of anthocyanin biosynthesis. BoMYB2 together with various BobHLHs specifically regulated the late anthocyanin biosynthetic pathway genes for anthocyanin biosynthesis. Our findings provide additional information for the complicated regulatory network of anthocyanin biosynthesis and the transcriptional regulation of transcription factors in vegetable crops.

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Abbreviations

PAL:

Phenylalanine ammonia lyase

C4H:

Cinnamate 4-hydroxylase

4CL:

4-Coumarate CoA ligase

CHS:

Chalcone synthase

CHI:

Chalcone isomerases

F3H:

Flavonone 3-hydroxylase

F3′H:

Flavonoid 3′-hydroxylase

DFR:

Dihydroflavonol reductase

LDOX:

Leucoanthocyanidin dioxygenase

UGT:

UDP glucose–flavonol glucosyl transferase

ACT:

Acyltransferase

OMT:

O-malonyltransferase

TTG1:

Transparent Testa Glabra 1

TT8:

Transparent Testa 8

TT2:

Transparent Testa 2

PAP1:

Production of Anthocyanin Pigment 1

PAP2:

Production of Anthocyanin Pigment 2

GL3:

Glabra 3

EGL3:

Enhancer of Glabra 3

bHLH:

Basic Helix-Loop-Helix

MBW:

MYB–bHLH–WD40

JAZ:

Jasmonate ZIM-domain

SPL:

Squamosa Promoter-Binding Protein-Like

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Acknowledgments

We thank Dr. Ted Thannhauser and Dr. Joyce Van Eck for critical review of this manuscript. We also thank Ms. Katherine Owsiany, Ms. Chrissa McFarlane and Ms. Veronica Kim for their technical support. This work was supported by the USDA-ARS base fund. USDA is an equal opportunity provider and employer. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture.

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  1. Li-Wei Chiu

    Present address: Pioneer Hi-Bred International, Inc., 4010 Point Eden Way, Hayward, CA, 94545, USA

Authors and Affiliations

  1. Department of Plant Breeding and Genetics, Cornell University, Ithaca, NY, 14853, USA

    Li-Wei Chiu & Li Li

  2. Robert W. Holley Center for Agriculture and Health, USDA-ARS, Cornell University, Ithaca, NY, 14853, USA

    Li Li

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  1. Li-Wei Chiu
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Correspondence to Li Li.

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Chiu, LW., Li, L. Characterization of the regulatory network of BoMYB2 in controlling anthocyanin biosynthesis in purple cauliflower. Planta 236, 1153–1164 (2012). https://doi.org/10.1007/s00425-012-1665-3

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