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An oilseed rape WRKY-type transcription factor regulates ROS accumulation and leaf senescence in Nicotiana benthamiana and Arabidopsis through modulating transcription of RbohD and RbohF

Abstract

Main conclusion

Overexpression of BnaWGR1 causes ROS accumulation and promotes leaf senescence. BnaWGR1 binds to promoters of RbohD and RbohF and regulates their expression.

Manipulation of leaf senescence process affects agricultural traits of crop plants, including biomass, seed yield and stress resistance. Since delayed leaf senescence usually enhances tolerance to multiple stresses, we analyzed the function of specific MAPK–WRKY cascades in abiotic and biotic stress tolerance as well as leaf senescence in oilseed rape (Brassica napus L.), one of the important oil crops. In the present study, we showed that expression of one WRKY gene from oilseed rape, BnaWGR1, induced an accumulation of reactive oxygen species (ROS), cell death and precocious leaf senescence both in Nicotiana benthamiana and transgenic Arabidopsis (Arabidopsis thaliana). BnaWGR1 regulates the transcription of two genes encoding key enzymes implicated in production of ROS, that is, respiratory burst oxidase homolog (Rboh) D and RbohF. A dual-luciferase reporter assay confirmed the transcriptional regulation of RbohD and RbohF by BnaWGR1. In vitro electrophoresis mobility shift assay (EMSA) showed that BnaWGR1 could bind to W-box cis-elements within promoters of RbohD and RbohF. Moreover, RbohD and RbohF were significantly upregulated in transgenic Arabidopsis overexpressing BnaWGR1. In summary, these results suggest that BnaWGR1 could positively regulate leaf senescence through regulating the expression of RbohD and RbohF genes.

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Abbreviations

CaMV:

Cauliflower mosaic virus

EMSA:

Electrophoretic mobility shift assay

GUS:

β-Glucuronidase

NBT:

Nitroblue tetrazolium

ROS:

Reactive oxygen species

SAG:

Senescence-associated gene

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Acknowledgements

This work was supported by the National Natural Science Foundation of China [No. 31301648 to BY and 31270293 to Y-QJ]. Natural Science Foundation of Shaanxi Province [no. 2014JQ3102 to BY]. The Fundamental Research Funds for the Central Universities of China [no. 2109021137, nos. 2452015413, 2452016009, to BY]. We thank Profs. Jian-Ye Chen (South China Agricultural University) and Benke Kuai (Fudan University) for technical assistance.

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Correspondence to Yuan-Qing Jiang or Bo Yang.

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Yang, L., Ye, C., Zhao, Y. et al. An oilseed rape WRKY-type transcription factor regulates ROS accumulation and leaf senescence in Nicotiana benthamiana and Arabidopsis through modulating transcription of RbohD and RbohF. Planta 247, 1323–1338 (2018). https://doi.org/10.1007/s00425-018-2868-z

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Keywords

  • Brassica napus
  • Leaf senescence
  • Rboh
  • ROS
  • Transcriptional regulation
  • WRKY