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The maize WRKY transcription factor ZmWRKY17 negatively regulates salt stress tolerance in transgenic Arabidopsis plants

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Abstract

Main conclusion

We cloned and characterized the ZmWRKY17 gene from maize. Overexpression of ZmWRKY17 in Arabidopsis led to increased sensitivity to salt stress and decreased ABA sensitivity through regulating the expression of some ABA- and stress-responsive genes.

The WRKY transcription factors have been reported to function as positive or negative regulators in many different biological processes including plant development, defense regulation and stress response. This study isolated a maize WRKY gene, ZmWRKY17, and characterized its role in tolerance to salt stress by generating transgenic Arabidopsis plants. Expression of the ZmWRKY17 was up-regulated by drought, salt and abscisic acid (ABA) treatments. ZmWRKY17 was localized in the nucleus with no transcriptional activation in yeast. Yeast one-hybrid assay showed that ZmWRKY17 can specifically bind to W-box, and it can activate W-box-dependent transcription in planta. Heterologous overexpression of ZmWRKY17 in Arabidopsis remarkably reduced plant tolerance to salt stress, as determined through physiological analyses of the cotyledons greening rate, root growth, relative electrical leakage and malondialdehyde content. Additionally, ZmWRKY17 transgenic plants showed decreased sensitivity to ABA during seed germination and early seedling growth. Transgenic plants accumulated higher content of ABA than wild-type (WT) plants under NaCl condition. Transcriptome and quantitative real-time PCR analyses revealed that some stress-related genes in transgenic seedlings showed lower expression level than that in the WT when treated with NaCl. Taken together, these results suggest that ZmWRKY17 may act as a negative regulator involved in the salt stress responses through ABA signalling.

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Abbreviations

MDA:

Malondialdehyde

REL:

Relative electrical leakage

TF:

Transcription factor

Tu:

Sodium tungstate

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Acknowledgements

We thank Chuanxiao Xie from Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing, China, for offering us the maize and Arabidopsis seeds in this study.

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Authors and Affiliations

  1. National Engineering Laboratory of Crop Stress Resistance Breeding, School of Life Sciences, Anhui Agricultural University, Hefei, 230036, China

    Ronghao Cai, Wei Dai, Congsheng Zhang, Yan Wang, Min Wu, Yang Zhao, Qing Ma, Yan Xiang & Beijiu Cheng

  2. Laboratory of Modern Biotechnology, School of Forestry and Landscape Architecture, Anhui Agricultural University, Hefei, 230036, China

    Yan Xiang

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  1. Ronghao Cai
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  2. Wei Dai
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  3. Congsheng Zhang
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  4. Yan Wang
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  9. Beijiu Cheng
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Correspondence to Yan Xiang or Beijiu Cheng.

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The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Funding

This work was supported by the National Key Research and Development Program (2017YFD0101205) and the Key Technologies Research and Development Program of Anhui Province (15czz03119).

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425_2017_2766_MOESM1_ESM.tif

Supplementary material 1 Fig. S1 Transcriptome analyses of WT and transgenic Arabidopsis plants overexpressing ZmWRKY17 gene. a The number of the significantly differentially expressed transcripts in WT and transgenic line (L10) after salt treatment. WT, wild type (untreated); WTNA, wild type treated with NaCl solution for 6 h; L10, transgenic line (untreated); L10NA, transgenic line treated with NaCl solution for 6 h. b Go analysis of wild Arabidopsis transcriptome after salt treatment in Solexa. c Go analysis of transgenic Arabidopsis transcriptome after salt treatment in Solexa (TIFF 10212 kb)

425_2017_2766_MOESM2_ESM.tif

Supplementary material 2 Fig. S2 Expression levels of stress-responsive genes in WT and two transgenic lines in response to salt stress. The induction of stress-responsive genes (RD29A, RD29B, RD22, KIN1, MYB101, NAC019 and NCED5) were measured by qRT-PCR analysis. Actin2 was used as the internal control. Data represent mean ± SD of three replicates (TIFF 9404 kb)

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Cai, R., Dai, W., Zhang, C. et al. The maize WRKY transcription factor ZmWRKY17 negatively regulates salt stress tolerance in transgenic Arabidopsis plants. Planta 246, 1215–1231 (2017). https://doi.org/10.1007/s00425-017-2766-9

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