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First published online February 8, 2008; 10.1104/pp.107.112789

Plant Physiology 146:1983-1995 (2008)
© 2008 American Society of Plant Biologists

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PLANTS INTERACTING WITH OTHER ORGANISMS

A Novel WRKY Transcription Factor Is Required for Induction of PR-1a Gene Expression by Salicylic Acid and Bacterial Elicitors[C],[W]

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

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

PR-1a is a salicylic acid-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::β-glucuronidase (GUS) genes in stably and transiently transformed tobacco indicated that NtWRKY12 and TGA1a act synergistically in PR-1a expression induced by salicylic acid and bacterial elicitors. Cotransfection of Arabidopsis thaliana 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.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Huub J.M. Linthorst (h.j.m.linthorst{at}biology.leidenuniv.nl).

[C] Some figures in this article are displayed in color online but in black and white in the print edition.

[W] The online version of this article contains Web-only data.

www.plantphysiol.org/cgi/doi/10.1104/pp.107.112789

* Corresponding author; e-mail h.j.m.linthorst{at}biology.leidenuniv.nl.

Received November 7, 2007; accepted January 31, 2008; published February 8, 2008.

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