Abstract

Rice stripe virus (RSV) causes rice stripe disease, which is one of the most serious rice diseases in eastern Asian countries. It has been shown that overexpression of RSV coat protein (CP) in rice plants enhances resistance against virus infection. However, the detailed mechanism underlying RSV CP-mediated virus resistance remains to be determined. In this study, we show that both translatable and non-translatable RSV CP transgenic Arabidopsis plants exhibited immunity to virus infection. By using deep sequencing analysis, transgene-derived small interfering RNAs (t-siRNAs) from non-translatable CP transgenic plants and virus-derived small interfering RNAs (vsiRNAs) mapping in the CP region from RSV-infected wild-type plants showed similar sequence distribution patterns, except for a significant increase in the abundance of t-siRNA reads compared with that of CP-derived vsiRNAs. To further test the correlation of t-siRNAs with RSV immunity, we developed RSV CP transgenic Arabidopsis plants in an siRNA-deficient dcl2/3/4 mutant background, and these CP transgenic plants showed the same sensitivity to RSV infection as non-transgenic plants. Together, our data indicate that the expression of RSV CP protein from a transgene is not a prerequisite for virus resistance and RSV CP-mediated resistance is mostly associated with the RNA silencing mechanism in Arabidopsis plants.

Highlights

  • RNA silencing is a conserved antiviral defense mechanism that has been used to increase resistance to plant virus infections (Pumplin and Voinnet, 2013; Guo et al, 2019)

  • By using deep sequencing analysis and comparing transgenicderived siRNAs (t-siRNAs) from non-translatable versions of coat protein (CP) transgenic plants and CP-derived virus-derived siRNAs from Rice stripe virus (RSV)-infected wild-type plants, we found that siRNA mapped to the CP sequence segment showed a similar pattern in both transgenic plants and virus-infected wild-type plants, except that greater accumulation of siRNAs was seen in transgenic immune plants

  • We provide evidence that the expression of RSV CP in transgenic Arabidopsis plants is not a prerequisite for CPmediated resistance

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Summary

Introduction

RNA silencing is a conserved antiviral defense mechanism that has been used to increase resistance to plant virus infections (Pumplin and Voinnet, 2013; Guo et al, 2019). Secondary vsiRNAs can be produced by dicing secondary viral dsRNA synthesized by host-encoded RNA-dependent RNA polymerases (RdRps) (Wassenegger and Krczal, 2006; Zhang et al, 2015). These vsiRNAs. RSV CP-Mediated Resistance are mainly loaded into the Argonaute (Ago) protein-containing RNA-induced silencing complex (RISC) to posttranscriptionally repress target viral and host RNAs by sequence complementarity (Carbonell and Carrington, 2015; Fang and Qi, 2016). In Arabidopsis plants, AGO1 and AGO2 are the two major plant antiviral argonautes against RNA viruses by associating with vsiRNAs based on the identity of the 5 terminal nucleotide (Mi et al, 2008; Carbonell and Carrington, 2015)

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