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DEAR1, a transcriptional repressor of DREB protein that mediates plant defense and freezing stress responses in Arabidopsis

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Abstract

Plants have evolved intricate mechanisms to respond and adapt to a wide variety of biotic and abiotic stresses in their environment. The Arabidopsis DEAR1 (DREB and EAR motif protein 1; At3g50260) gene encodes a protein containing significant homology to the DREB1/CBF (dehydration-responsive element binding protein 1/C-repeat binding factor) domain and the EAR (ethylene response factor-associated amphiphilic repression) motif. We show here that DEAR1 mRNA accumulates in response to both pathogen infection and cold treatment. Transgenic Arabidopsis overexpressing DEAR1 (DEAR1ox) showed a dwarf phenotype and lesion-like cell death, together with constitutive expression of PR genes and accumulation of salicylic acid. DEAR1ox also showed more limited P. syringae pathogen growth compared to wild-type, consistent with an activated defense phenotype. In addition, transient expression experiments revealed that the DEAR1 protein represses DRE/CRT (dehydration-responsive element/C-repeat)-dependent transcription, which is regulated by low temperature. Furthermore, the induction of DREB1/CBF family genes by cold treatment was suppressed in DEAR1ox, leading to a reduction in freezing tolerance. These results suggest that DEAR1 has an upstream regulatory role in mediating crosstalk between signaling pathways for biotic and abiotic stress responses.

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Acknowledgments

We are grateful to Dr. Shibuya for providing chitin oligomers, Ms. Fukazawa, Dr. Hayashi and Dr. Nishimura for microarray analysis, Ms. Yoko Osaka and Ms. Natsuko Yamoto for technical assistance, and Dr. Derek B. Goto for critical reading of the manuscript. This work was supported by a Grant-in-Aid for Scientific Research (nos. 19657013, 19039001) to J.Y., in part by the Program from Basic Research Activities for Innovative Bioscience (PROBRAIN), by grants from the 21st century COE Hokkaido University (J.Y., T.T.) and The Akiyama Foundation to J.Y.

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Authors and Affiliations

  1. Faculty of Advanced Life Science, Graduate School of Life Science, Hokkaido University, Kita-ku N10-W8, Sapporo, 060-0810, Japan

    Tomokazu Tsutsui, Wataru Kato, Yutaka Asada, Kaori Sako, Takeo Sato, Yutaka Sonoda, Akira Ikeda & Junji Yamaguchi

  2. Laboratory of Plant Molecular Physiology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo, 113-8657, Japan

    Satoshi Kidokoro & Kazuko Yamaguchi-Shinozaki

  3. Biological Resources Division, Japan International Research Center for Agricultural Sciences, Tsukuba, Ibaraki, 305-8686, Japan

    Kazuko Yamaguchi-Shinozaki

  4. Biodiversity Conservation Research Project, National Institute for Environmental Studies, 16-2 Onogawa, Tsukuba, Ibaraki, 305-8506, Japan

    Masanori Tamaoki

  5. Graduate School of Agriculture, Hokkaido University, Kita-ku N9-W9, Sapporo, 060-8589, Japan

    Keita Arakawa

  6. Plant Functional Genomics Research Group, RIKEN Plant Science Center (PSC), RIKEN, Yokohama Institute, Yokohama, Kanagawa, 230-0045, Japan

    Takanari Ichikawa, Miki Nakazawa, Kazuo Shinozaki & Minami Matsui

  7. Plant Genomic Network Research Group, RIKEN Plant Science Center (PSC), RIKEN, Yokohama Institute, Yokohama, Kanagawa, 230-0045, Japan

    Motoaki Seki

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  1. Tomokazu Tsutsui
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Correspondence to Junji Yamaguchi.

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T. Tsutsui, W. Kato and A. Ikeda contributed equally to this work.

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Tables S1 and S2 (DOC 80 kb)

10265_2009_252_MOESM2_ESM.tif

Fig. S1. Phylogenetic Tree of DEAR Family. A multiple-sequence alignment constructed with Clustal W is displayed using Tree View (Morita-Yamamuro et al. 2005). Sequences were obtained from GenBank, and accession numbers are as follows: Arabidopsis thaliana (DEAR1), Q9SNE1; Arabidopsis thaliana (DEAR2), Q9FH94; Arabidopsis thaliana (DEAR3), O22174; Arabidopsis thaliana (DEAR4), ABF83647; Arabidopsis thaliana (DEAR5), ABD57514; Arabidopsis thaliana (DEAR6), ABD57516; Oryza sativa (Os04g0648900), NP_001054079; Oryza sativa (Os06g0166400), NP_001056914; Triticum aestivum, AAY98505; Vitis vinifera, AAZ74793; Vitis vinifera, CAO61384; Gossypium hirsutum, AAO43165; Glycine max, ABB36645; Glycine max, AAP47161. The scale represents 0.1 substitutions per site. (TIFF 67 kb)

10265_2009_252_MOESM3_ESM.tif

Fig. S2. Characterization of 35S::DEAR1 and DEAR1ox Transgenic Lines.(a) Morphology of 4-week-old wild-type Col-0 (WT), 35S::DEAR1 and DEAR1ox plants. Bar = 1 cm.(b) Overexpression of DEAR1 gene in 35S::DEAR1 and DEAR1ox plants. Total RNA from 4-week-old vegetative shoot tissue of wild-type (WT), 35S::DEAR1 and DEAR1ox was analyzed by RT-PCR. EF1α was used as a control.(c) Imaging DEAR1::GFP in DEAR1ox. Epi-fluorescence microscope images by Bright field and GFP epi-fluorescence images of root from the transgenic plant for the 35S::GFP and DEAR1ox. Bar = 200 μm. (TIFF 1044 kb)

10265_2009_252_MOESM4_ESM.tif

Fig. S3. Expression Profiles of DEAR1 in Response to Various Abiotic Stresses.Expression profiles of DEAR1 in wild-type plants during cold (closed circle; sky blue), osmotic (closed square; yellow), salt (closed triangle; light blue), drought (asterisk; red), oxidative (open diamond; light green), UV-B (open circle; light purple), wounding (open square; aquamarine) and heat (open triangle; orange) stresses. No stress exposure is as also shown as a control (closed diamond; black). Data was compiled from microarray data available in the Arabidopsis eFP Browser (http://bar.utoronto.ca/efp/cgi-bin/efpWeb.cgi). (TIFF 344 kb)

10265_2009_252_MOESM5_ESM.tif

Fig. S4. Expression Profiles of DEAR1 in Response to Pathogen Infection.(a) Expression of DEAR1 in wild-type plants following infection with P. syringae pv. tomato DC3000 (closed square; orange) and 10 mM MgCl2 as a mock control (closed diamond; black). Data was compiled from microarray data available in the Arabidopsis eFP Browser (http://bar.utoronto.ca/efp/cgi-bin/efpWeb.cgi).(b) Expression of DEAR1 in wild-type plants following infection with B. cinerea (closed square; orange) and potato dextrose broth as a mock control (closed diamond; black). Data was compiled from microarray data available in the Arabidopsis eFP Browser (http://bar.utoronto.ca/efp/cgi-bin/efpWeb.cgi). (TIFF 256 kb)

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Tsutsui, T., Kato, W., Asada, Y. et al. DEAR1, a transcriptional repressor of DREB protein that mediates plant defense and freezing stress responses in Arabidopsis. J Plant Res 122, 633–643 (2009). https://doi.org/10.1007/s10265-009-0252-6

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