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Promoter analysis reveals cis-regulatory motifs associated with the expression of the WRKY transcription factor CrWRKY1 in Catharanthus roseus

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Abstract

WRKY transcription factors (TFs) are emerging as an important group of regulators of plant secondary metabolism. However, the cis-regulatory elements associated with their regulation have not been well characterized. We have previously demonstrated that CrWRKY1, a member of subgroup III of the WRKY TF family, regulates biosynthesis of terpenoid indole alkaloids in the ornamental and medicinal plant, Catharanthus roseus. Here, we report the isolation and functional characterization of the CrWRKY1 promoter. In silico analysis of the promoter sequence reveals the presence of several potential TF binding motifs, indicating the involvement of additional TFs in the regulation of the TIA pathway. The CrWRKY1 promoter can drive the expression of a β-glucuronidase (GUS) reporter gene in native (C. roseus protoplasts and transgenic hairy roots) and heterologous (transgenic tobacco seedlings) systems. Analysis of 5′- or 3′-end deletions indicates that the sequence located between positions −140 to −93 bp and −3 to +113 bp, relative to the transcription start site, is critical for promoter activity. Mutation analysis shows that two overlapping as-1 elements and a CT-rich motif contribute significantly to promoter activity. The CrWRKY1 promoter is induced in response to methyl jasmonate (MJ) treatment and the promoter region between −230 and −93 bp contains a putative MJ-responsive element. The CrWRKY1 promoter can potentially be used as a tool to isolate novel TFs involved in the regulation of the TIA pathway.

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Abbreviations

as-1 :

Activation sequence 1

MJ:

Methyl jasmonate

TIA:

Terpenoid indole alkaloids

TSS:

Transcription start site

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Acknowledgments

ZY acknowledges the receipt of a pre-doctoral fellowship from Shanxi Agriculture University, China.

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Authors and Affiliations

  1. Institute of Molecular Agriculture and Bioenergy, Shanxi Agricultural University, Taigu, 030801, Shanxi, China

    Zhirong Yang & Runzhi Li

  2. Department of Biochemistry, College of Arts and Sciences, Shanxi Agricultural University, Taigu, 030801, Shanxi, China

    Zhirong Yang

  3. Department of Plant and Soil Sciences, Kentucky Tobacco Research and Development Center, University of Kentucky, Lexington, KY, 40546, USA

    Barunava Patra, Sitakanta Pattanaik & Ling Yuan

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  1. Zhirong Yang
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  2. Barunava Patra
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  3. Runzhi Li
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  4. Sitakanta Pattanaik
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  5. Ling Yuan
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Corresponding authors

Correspondence to Sitakanta Pattanaik or Ling Yuan.

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Z. Yang and B. Patra contributed equally to this work.

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Yang, Z., Patra, B., Li, R. et al. Promoter analysis reveals cis-regulatory motifs associated with the expression of the WRKY transcription factor CrWRKY1 in Catharanthus roseus . Planta 238, 1039–1049 (2013). https://doi.org/10.1007/s00425-013-1949-2

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