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ROS accumulation and antiviral defence control by microRNA528 in rice

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Abstract

MicroRNAs (miRNAs) are key regulators of plant–pathogen interactions. Modulating miRNA function has emerged as a new strategy to produce virus resistance traits15. However, the miRNAs involved in antiviral defence and the underlying mechanisms remain largely elusive. We previously demonstrated that sequestration by Argonaute (AGO) proteins plays an important role in regulating miRNA function in antiviral defence pathways6. Here we reveal that cleavage-defective AGO18 complexes sequester microRNA528 (miR528) upon viral infection. We show that miR528 negatively regulates viral resistance in rice by cleaving L-ascorbate oxidase (AO) messenger RNA, thereby reducing AO-mediated accumulation of reactive oxygen species. Upon viral infection, miR528 becomes preferentially associated with AGO18, leading to elevated AO activity, higher basal reactive oxygen species accumulation and enhanced antiviral defence. Our findings reveal a mechanism in which antiviral defence is boosted through suppression of an miRNA that negatively regulates viral resistance. This mechanism could be manipulated to engineer virus-resistant crop plants.

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Figure 1: MiR528 is required for antiviral immunity.
Figure 2: Transgenic expression of miR528-resistant AO mRNA-enhanced virus resistance in rice.
Figure 3: AO enhanced accumulation of ROS in rice.
Figure 4: A model of miRNA528 function in regulating plant antiviral immunity.

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Acknowledgements

We thank F. Qu (Ohio State University) and Y. Li (Tsinghua-Peking Center for Life Sciences) for critical reading of the manuscript, L. Li (Peking University) for technical assistance, G. Liu (Tsinghua University) for assistance with the AO and AsA assay and the Integrated R & D Services – WuXi AppTec for generating the antisera used in this study. This work was supported by grants from the National Basic Research Program 973 (2014CB138400), Natural Science Foundation of China (91540203, 31530062, 31420103904, 31123007 and 31272018), the National Key Research and Development Program of China (2016YFD0100904), the Transgenic Research Program (2016ZX08010-001 and 2016ZX08009001-005) and state key laboratories of protein and plant gene research, plant genomics. J.W. was supported in part by the Postdoctoral Fellowship of Peking-Tsinghua Center for Life Sciences.

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J.W., Z.Y., X.C. and Y. Li designed the experiments; J.W., Z.Y., R.Y., Z.Y., S.Z., Y.W., L.J., P.L., X.S., T.Z. and Y. Lan performed the experiments; J.W., Z.Y., R.Y., L.X., X.Z., C.C., Y.Q., X.C. and Y. Li analysed the data; J.W., Z.Y. and Y. Li wrote the paper. All the authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Xiaofeng Cao or Yi Li.

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The authors declare no competing financial interests.

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Supplementary Figures 1–10, Supplementary Tables 1–3. (PDF 3498 kb)

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Wu, J., Yang, R., Yang, Z. et al. ROS accumulation and antiviral defence control by microRNA528 in rice. Nature Plants 3, 16203 (2017). https://doi.org/10.1038/nplants.2016.203

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