title: Promoter analysis reveals cis-regulatory motifs associated with the expression of the WRKY transcription factor CrWRKY1 in Catharanthus roseus.
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.
authors: Institute of Molecular Agriculture and Bioenergy, Shanxi Agricultural University, Taigu, 030801, Shanxi, China.|Department of Biochemistry, College of Arts and Sciences, Shanxi Agricultural University, Taigu, 030801, Shanxi, China.
authors: Zhirong
authors: Z
authors: Yang
authors: Department of Plant and Soil Sciences, Kentucky Tobacco Research and Development Center, University of Kentucky, Lexington, KY, 40546, USA.
authors: Barunava
authors: B
authors: Patra
authors: Institute of Molecular Agriculture and Bioenergy, Shanxi Agricultural University, Taigu, 030801, Shanxi, China.
authors: Runzhi
authors: R
authors: Li
authors: Department of Plant and Soil Sciences, Kentucky Tobacco Research and Development Center, University of Kentucky, Lexington, KY, 40546, USA. spatt2@uky.edu.
authors: Sitakanta
authors: S
authors: Pattanaik
authors: Department of Plant and Soil Sciences, Kentucky Tobacco Research and Development Center, University of Kentucky, Lexington, KY, 40546, USA. lyuan3@uky.edu.
authors: Ling
authors: L
authors: Yuan
mesh_terms: D000085:Acetates / Q000494:pharmacology; D030008:Catharanthus / Q000235:genetics* / Q000378:metabolism; D003517:Cyclopentanes / Q000494:pharmacology; D018744:DNA, Plant / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017343:Genes, Plant; D017930:Genes, Reporter; D053858:Metabolic Networks and Pathways; D054883:Oxylipins / Q000494:pharmacology; D010937:Plant Growth Regulators / Q000494:pharmacology; D010940:Plant Proteins / Q000235:genetics*; D018517:Plant Roots / Q000235:genetics; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000187:drug effects; D046948:Secologanin Tryptamine Alkaloids / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*; D024363:Transcription Initiation Site
chemical_list: D000085:Acetates; D003517:Cyclopentanes; D018744:DNA, Plant; D054883:Oxylipins; D010937:Plant Growth Regulators; D010940:Plant Proteins; D046948:Secologanin Tryptamine Alkaloids; D014157:Transcription Factors; C072239:methyl jasmonate
keywords: Catharanthus roseus; Hairy roots; Terpenoid indole alkaloids; WRKY transcription factor
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