Functional analysis of a WRKY transcription factor involved in transcriptional activation of the DBAT gene in Taxus chinensis. (12th June 2012)

• Record Type: Journal Article
• Authors: Li, S.; Zhang, P.; Zhang, M.; Fu, C.; Yu, L.
• Is Part Of: Plant biology. Volume 15:Number 1(2013:Jan.)
• Page Start: 19
• Page End: 26
• Publication Date: 2012-06-12
• Subjects: Botany -- Periodicals; Plants -- genetics -- Periodicals; Plants -- growth & development -- Periodicals; Plant Proteins -- Periodicals; Gene Expression Regulation, Plant -- Periodicals; Botanique -- Périodiques; 580
• DOI: 10.1111/j.1438-8677.2012.00611.x
• Languages: English
• ISSNs: 1435-8603

Abstract

<abstract abstract-type="main" xml:lang="en"> <title>Abstract</title> <p>Although the regulation of taxol biosynthesis at the transcriptional level remains unclear, 10‐deacetylbaccatin III‐10 β‐<italic>O</italic>‐acetyl transferase (DBAT) is a critical enzyme in the biosynthesis of taxol. The 1740 bp fragment 5′‐flanking sequence of the <italic>dbat</italic> gene was cloned from <italic>Taxus chinensis</italic> cells. Important regulatory elements needed for activity of the <italic>dbat</italic> promoter were located by deletion analyses in <italic>T. chinensis</italic> cells. A novel WRKY transcription factor, <italic>TcWRKY1</italic>, was isolated with the yeast one‐hybrid system from a <italic>T. chinensis</italic> cell cDNA library using the important regulatory elements as bait. The gene expression of <italic>TcWRKY1</italic> in <italic>T. chinensis</italic> suspension cells was specifically induced by methyl jasmonate (MeJA). Biochemical analysis indicated that <italic>TcWRKY1</italic> protein specifically interacts with the two W‐box (TGAC) <italic>cis</italic>‐elements among the important regulatory elements. Overexpression of <italic>TcWRKY1</italic> enhanced <italic>dbat</italic> expression in <italic>T. chinensis</italic> suspension cells, and RNA interference (RNAi) reduced the level of transcripts of <italic>dbat</italic>. These results suggest that <italic>TcWRKY1</italic> participates in regulation of taxol biosynthesis in <italic>T. chinensis</italic> cells, and that <italic>dbat</italic> is a target gene of this transcription factor. This research also provides a potential candidate gene for engineering increased taxol accumulation in <italic>Taxus</italic> cell cultures.</p> </abstract>