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Phylogenetic analysis of barley WRKY proteins and characterization of HvWRKY1 and -2 as repressors of the pathogen-inducible gene HvGER4c

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Abstract

The WRKY transcription factors belong to an evolutionarily conserved superprotein family predominantly present in the plant kingdom. WRKY proteins of barley are not yet fully annotated and most of them are not functionally characterized. We performed a genome-wide identification of WRKY members based on the recently accessible barley draft genome sequence and full-length cDNA datasets. As a result, 34 novel putative proteins have been identified which extend the existing list for barley WRKYs to 94. Phylogenetic analysis of the WRKY domains allowed ranking into three groups (I, II, III), with an expansion in group III in monocots. Two members of subgroup IIa, the wound and pathogen-inducible HvWRKY1 and HvWRKY2, are known as negative defense regulators. Here, we demonstrate that both transcription factors repress the activity of the powdery mildew-induced promoter of HvGER4c, a germin-like defense-related protein. The repression did not require the negative defense regulator MLO nor was it affected by the presence of the R protein MLA12. Moreover, the expression of the Arabidopsis ortholog AtWRKY40 in barley compromised basal resistance to powdery mildew, providing evidence for functional conservation of sequence-related WRKY proteins across monocots and dicots.

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Acknowledgments

We thank A. Himmelbach and P. Schweizer for providing the series of pHvGER4c::GUS plasmids and I.E. Somssich for providing the pDONR-AtWRKY40 plasmid. A. Zuccaro is acknowledged for help with CLSM and critical reading of the manuscript. This work was supported by the Bundesministerium für Bildung und Forschung Grant 0313124 and a Justus Liebig University Giessen PhD scholarship to D.L.

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The authors declare that they have no conflict of interest.

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  1. Dilin Liu

    Present address: Rice Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, 510640, China

  2. Puyan Zhao

    Present address: College of Horticulture, South China Agricultural University, Guangzhou, 510642, China

Authors and Affiliations

  1. Research Centre for BioSystems, Land Use, and Nutrition (IFZ Giessen), Institute of Phytopathology and Applied Zoology, Justus Liebig University Giessen, 35392, Giessen, Germany

    Dilin Liu, Katja Leib, Puyan Zhao, Karl-Heinz Kogel & Gregor Langen

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  1. Dilin Liu
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Correspondence to Gregor Langen.

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Liu, D., Leib, K., Zhao, P. et al. Phylogenetic analysis of barley WRKY proteins and characterization of HvWRKY1 and -2 as repressors of the pathogen-inducible gene HvGER4c . Mol Genet Genomics 289, 1331–1345 (2014). https://doi.org/10.1007/s00438-014-0893-6

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