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Root-specific CLE3 expression is required for WRKY33 activation in Arabidopsis shoots

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Abstract

Key message

This study focused on the role of CLE1–7 peptides as defense mediators, and showed that root-expressed CLE3 functions as a systemic signal to regulate defense-related gene expression in shoots.

Abstract

In the natural environment, plants employ diverse signaling molecules including peptides to defend themselves against various pathogen attacks. In this study, we investigated whether CLAVATA3/EMBRYO SURROUNDING REGION-RELATED (CLE) genes (CLE1–7) respond to biotic stimuli. CLE3 showed significant up-regulation upon treatment with flg22, Pep2, and salicylic acid (SA). Quantitative real-time PCR (qRT-PCR) analysis revealed that CLE3 expression is regulated by the NON-EXPRESSOR OF PR GENES1 (NPR1)-dependent SA signaling and flg22–FLAGELLIN-SENSITIVE 2 (FLS2) signaling pathways. We demonstrated that SA-induced up-regulation of CLE3 in roots was required for activation of WRKY33, a gene involved in the regulation of systemic acquired resistance (SAR), in shoots, suggesting that CLE3 functions as a root-derived signal that regulates the expression of defense-related genes in shoots. Microarray analysis of transgenic Arabidopsis lines overexpressing CLE3 under the control of a β-estradiol-inducible promoter revealed that root-confined CLE3 overexpression affected gene expression in both roots and shoots. Comparison of CLE2- and CLE3-induced genes indicated that CLE2 and CLE3 peptides target a few common but largely distinct downstream genes. These results suggest that root-derived CLE3 is involved in the regulation of systemic rather than local immune responses. Our study also sheds light on the potential role of CLE peptides in long-distance regulation of plant immunity.

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Data Availability

Microarray data generated in this study were deposited to the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) database under the accession number GSE176064. Web link; https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE176064.

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Acknowledgements

We thank Kyoko Ohashi-Ito and Taku Demura for providing the transgenic plants used in this study and pBGYN vector, respectively. We appreciate Yusuke Saijo, Kenichi Tsuda and Imre Somsich for sharing the seeds used in this study. We also thank Yukiko Sugisawa, Yumi Suzuki, Masakazu Ota, and Kuninori Iwamoto for technical assistance.

Funding

This work was supported in part by Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology (15H05958 to HF); Japan Society for the Promotion of Science (21H02500 to HF); PRESTO (117665 to SB) and ERATO (JPMJER1502 to SB) from Japan Science and Technology Agency; and JPNP18016 (to SB) commissioned by New Energy and Industrial Technology Development Organization (NEDO).

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Author notes

  1. Dichao Ma and Satoshi Endo have contributed equally to this work.

Authors and Affiliations

  1. Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan

    Dichao Ma, Satoshi Endo & Hiroo Fukuda

  2. Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan

    Dichao Ma & Toru Fujiwara

  3. Institute of Interdisciplinary Research, Kyoto University of Advanced Science, 1-1 Nanjo-Ohtani, Sogabe-cho, Kameoka-city, Kyoto, 621-8555, Japan

    Satoshi Endo

  4. Department of Plant Life Science, Faculty of Agriculture, Ryukoku University, 1-5 Yokotani, Seta Oe-cho, Otsu, Shiga, 520-2194, Japan

    Eriko Betsuyaku & Shigeyuki Betsuyaku

  5. Department of Bioscience and Biotechnology, Faculty of Bioenvironmental Sciences, Kyoto University of Advanced Science, 1-1 Nanjo-Ohtani, Sogabe-cho, Kameoka-city, Kyoto, 621-8555, Japan

    Hiroo Fukuda

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  1. Dichao Ma
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Contributions

DM, SE, EB, SB, and HF designed the research; DM, SE, EB, and SB performed the experiments; and DM, SE, TF, SB, and HF wrote the manuscript.

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Correspondence to Shigeyuki Betsuyaku or Hiroo Fukuda.

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Ma, D., Endo, S., Betsuyaku, E. et al. Root-specific CLE3 expression is required for WRKY33 activation in Arabidopsis shoots. Plant Mol Biol 108, 225–239 (2022). https://doi.org/10.1007/s11103-021-01234-9

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  • DOI: https://doi.org/10.1007/s11103-021-01234-9

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