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Differential combinatorial interactions of cis-acting elements recognized by R2R3-MYB, BZIP, and BHLH factors control light-responsive and tissue-specific activation of phenylpropanoid biosynthesis genes

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Abstract

Chalcone synthase (CHS), chalcone flavanone isomerase (CFI), flavanone 3-hydroxylase (F3H) and flavonol synthase (FLS) catalyze successive steps in the biosynthetic pathway leading to the production of flavonols. We show that in Arabidopsis thaliana all four corresponding genes are coordinately expressed in response to light, and are spatially coexpressed in siliques, flowers and leaves. Light regulatory units (LRUs) sufficient for light responsiveness were identified in all four promoters. Each unit consists of two necessary elements, namely a MYB-recognition element (MRE) and an ACGT-containing element (ACE). C1 and Sn, a R2R3-MYB and a BHLH factor, respectively, known to control tissue specific anthocyanin biosynthesis in Z. mays, were together able to activate the AtCHS promoter. This activation of the CHS promoter required an intact MRE and a newly identified sequence designated R response element (RRE AtCHS) containing the BHLH factor consensus binding site CANNTG. The RRE was dispensable for light responsiveness, and the ACE was not necessary for activation by C1/Sn. These data suggest that a BHLH and a R2R3-MYB factor cooperate in directing tissue-specific production of flavonoids, while an ACE-binding factor, potentially a BZIP, and a R2R3-MYB factor work together in conferring light responsiveness.

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Abbreviations

ACE:

ACGT-containing element

ACS:

ACE-like sequence

BHLH:

basic helix-loop-helix

BZIP:

basic region/leucine zipper

CFI:

chalcone flavanone isomerase

CHS:

chalcone synthase

chx:

cycloheximide

EMSA:

electrophoretic mobility shift assay

F3H:

flavanone 3-hydroxylase

FLS:

flavonol synthase

LRU:

light regulatory unit

MRE:

MYB-recognition element

MRS:

MRE-like sequence

RRE:

R response element

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Authors and Affiliations

  1. Department of Plant Breeding and Yield Physiology, Max-Planck-Institute for Plant Breeding Research, Carl-von-Linné-Weg 10, D-50829, Köln, Germany

    Ulrike Hartmann, Martin Sagasser, Frank Mehrtens, Ralf Stracke & Bernd Weisshaar

  2. Genome Research, Department of Biology, Bielefeld University, D-33594, Bielefeld, Germany

    Martin Sagasser, Frank Mehrtens, Ralf Stracke & Bernd Weisshaar

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  1. Ulrike Hartmann
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  2. Martin Sagasser
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  3. Frank Mehrtens
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  4. Ralf Stracke
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  5. Bernd Weisshaar
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Correspondence to Bernd Weisshaar.

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Hartmann, U., Sagasser, M., Mehrtens, F. et al. Differential combinatorial interactions of cis-acting elements recognized by R2R3-MYB, BZIP, and BHLH factors control light-responsive and tissue-specific activation of phenylpropanoid biosynthesis genes. Plant Mol Biol 57, 155–171 (2005). https://doi.org/10.1007/s11103-004-6910-0

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