Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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Published on February 8, 2008; 10.1104/pp.107.112789

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Received November 7, 2007
Accepted January 31, 2008

A NOVEL WRKY TRANSCRIPTION FACTOR IS REQUIRED FOR INDUCTION OF PR-1A GENE EXPRESSION BY SALICYLIC ACID AND BACTERIAL ELICITORS

Marcel C. van Verk , Dimitri Pappaioannou , Lyda Neeleman , John F. Bol , and Huub J.M. Linthorst *

Institute of Biology, Leiden University, Clusius Laboratory, Wassenaarseweg 64, 2333 AL Leiden, The Netherlands

* Corresponding author; email: h.j.m.linthorst{at}biology.leidenuniv.nl.

PR-1a is a salicylic acid (SA)-inducible defense gene of tobacco (Nicotiana tabacum). One-hybrid screens identified a novel tobacco WRKY-transcription factor (NtWRKY12) with specific binding sites in the PR-1a promoter at positions -564 (box WK1) and -859 (box WK2). NtWRKY12 belongs to the class of transcription factors in which the WRKY sequence is followed by a GKK rather than a GQK sequence. The binding sequence of NtWRKY12 (WK box TTTTCCAC) deviated significantly from the consensus sequence (W box TTGAC[C/T]) shown to be recognized by WRKY factors with the GQK sequence. Mutation of the GKK sequence in NtWRKY12 into GQK or GEK abolished binding to the WK box. The WK1 box is in close proximity to binding sites in the PR-1a promoter for transcription factors TGA1a (as-1 box) and Myb1 (MBSII box). Expression studies with PR-1a promoter::GUS genes in stably and transiently transformed tobacco indicated that NtWRKY12 and TGA1a act synergistically in PR-1a expression induced by SA and bacterial elicitors. Co-transfection of Arabidopsis protoplasts with 35S::NtWRKY12 and PR-1a::GUS promoter fusions showed that overexpression of NtWRKY12 resulted in a strong increase in GUS expression, which required functional WK boxes in the PR-1a promoter.




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