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Identification and characterisation of a glycine-rich RNA-binding protein as an endogenous suppressor of RNA silencing from Nicotiana glutinosa

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Abstract

Main conclusion

This study shows that NgRBP suppresses both local and systemic RNA silencing induced by sense- or double-stranded RNA, and the RNA binding activity is essential for its function.

To counteract host defence, many plant viruses encode viral suppressors of RNA silencing targeting various stages of RNA silencing. There is increasing evidence that the plants also encode endogenous suppressors of RNA silencing (ESR) to regulate this pathway. In this study, using Agrobacterium infiltration assays, we characterized NgRBP, a glycine-rich RNA-binding protein from Nicotiana glutinosa, as an ESR. Our results indicated that NgRBP suppressed both local and systemic RNA silencing induced by sense- or double-stranded RNA. We also demonstrated that NgRBP could promote Potato Virus X (PVX) infection in N. benthamiana. NgRBP knockdown by virus-induced gene silencing enhanced PVX and Cucumber mosaic virus resistance in N. glutinosa. RNA immunoprecipitation and electrophoretic mobility shift assays showed that NgRBP bound to GFP mRNA, dsRNA rather than siRNA. These findings provide the evidence that NgRBP acts as an ESR and the RNA affinity of NgRBP plays the key role in its ESR activity. NgRBP responds to multiple signals such as ABA, MeJA, SA, and Tobacco mosaic virus infection. Therefore, it could participate in the regulation of gene expression under specific conditions.

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Abbreviations

CMV:

Cucumber mosaic virus

EMSA:

Electrophoretic mobility shift assay

ESR:

Endogenous suppressor of RNA silencing

GD:

Glycine-rich domain

GRP:

Glycine-rich RNA-binding protein

PVX:

Potato virus X

RBP:

RNA-binding protein

RIP:

RNA immunoprecipitation

RRM:

RNA recognition motif

RSS:

RNA-silencing suppressor

TMV:

Tobacco mosaic virus

VSR:

Viral suppressor of RNA silencing

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Acknowledgements

We thank Dr. David Baulcombe for providing P19, PVX vector, and N. benthamiana line 16c seeds. This work was supported by Shandong Provincial Natural Science Foundation, China [ZR2015CM018, ZR2018LC009], Genetically Modified Organisms Breeding Major Projects of China [2016ZX08001-002], Shandong Medical and Health Science and Technology Development Plan Project [2017WS426].

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  1. Xu Huang and Ru Yu contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Tai′an, Shandong, China

    Xu Huang, Ru Yu, Wenjing Li, Liwei Geng, Changxiang Zhu & Hongmei Liu

  2. Institute of Immunology, Taishan Medical University, Tai′an, Shandong, China

    Xiuli Jing

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  1. Xu Huang
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  2. Ru Yu
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  3. Wenjing Li
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Correspondence to Hongmei Liu.

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Huang, X., Yu, R., Li, W. et al. Identification and characterisation of a glycine-rich RNA-binding protein as an endogenous suppressor of RNA silencing from Nicotiana glutinosa. Planta 249, 1811–1822 (2019). https://doi.org/10.1007/s00425-019-03122-5

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