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The rice transcription factor OsWRKY47 is a positive regulator of the response to water deficit stress

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Abstract

OsWRKY47 is a divergent rice transcription factor belonging to the group II of the WRKY family. A transcriptomic analysis of the drought response of transgenic rice plants expressing P SARK ::IPT, validated by qPCR, indicated that OsWRKY47 expression was induced under drought stress in P SARK ::IPT plants. A PCR-assisted site selection assay (SELEX) of recombinant OsWRKY47 protein showed that the preferred sequence bound in vitro is (G/T)TTGACT. Bioinformatics analyses identified a number of gene targets of OsWRKY47; among these two genes encode a Calmodulin binding protein and a Cys-rich secretory protein. Using Oswrk47 knockout mutants and transgenic rice overexpressing OsWRKY47 we show that the transcription of these putative targets were regulated by OsWRKY47. Phenotypic analysis carried out with transgenic rice plants showed that Oswrky47 mutants displayed higher sensitivity to drought and reduced yield, while plants overexpressing OsWRKY47 were more tolerant.

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Acknowledgments

This research was supported by funds from the UC Office of the President, UC Discovery Grant # 212976 (EB). The research of the Argentinean group was supported by Agencia Nacional de Promoción Científica y Tecnológica (PICT 2011 0850 and PICT 2012 0955) and Universidad Nacional del Litoral (UNL). JR is a CONICET Ph. D. Fellow and RLC is a career member of the same institution.

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    Authors and Affiliations

    1. Instituto de Agrobiotecnología del Litoral (CONICET-UNL), Colectora ruta 168 km 0, Paraje El Pozo, 3000, Santa Fe, Argentina

      Jesica Raineri & Raquel Lia Chan

    2. Department of Plant Sciences – Mail Stop 5, University of California, 1 Shields Ave, Davis, CA, 95616, USA

      Songhu Wang, Zvi Peleg & Eduardo Blumwald

    3. Chengdu Institute of Biology, Chinese Academy of Science, Chengdu, Sichuan, China

      Songhu Wang

    4. The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, The Hebrew University of Jerusalem, P.O. Box 12, 7610001, Rehovot, Israel

      Zvi Peleg

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    1. Jesica Raineri
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    2. Songhu Wang
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    3. Zvi Peleg
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    4. Eduardo Blumwald
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    5. Raquel Lia Chan
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    Corresponding author

    Correspondence to Eduardo Blumwald.

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    Jesica Raineri and Songhu Wang have contributed equally to this work.

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    Fig. S1.

    Schematic representation of the in silico analysis performed to detect putative targets of OsWRKY47. Supplementary material 1 (JPEG 38 kb)

    Fig. S2.

    Validation of the expression of candidate genes targeted by OsWRKY47 by qRT-PCR. Supplementary material 2 (PDF 115 kb)

    Table S1.

    List of genes containing OsWRKY47-bound sequences in their promoters. Supplementary material 3 (XLSX 14 kb)

    Table S2.

    List of genes co-expressed with OsWRKY47 according to in silico analysis. Supplementary material 4 (XLSX 12 kb)

    Table S3.

    List of oligonucleotides used in this work for cloning or for RT-qPCR. Supplementary material 5 (DOCX 23 kb)

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    Raineri, J., Wang, S., Peleg, Z. et al. The rice transcription factor OsWRKY47 is a positive regulator of the response to water deficit stress. Plant Mol Biol 88, 401–413 (2015). https://doi.org/10.1007/s11103-015-0329-7

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