水稻条纹病毒 NS3 蛋白影响本氏烟内源基因的表达。
NS3 Protein from Rice stripe virus affects the expression of endogenous genes in Nicotiana benthamiana.
机构信息
Chongqing Key Laboratory of Plant Disease Biology, College of Plant Protection, Southwest University, Chongqing, 400716, China.
The State Key Laboratory Breading Base for Sustainable Control of Pest and Disease, Key Laboratory of Biotechnology in Plant Protection of MOA of China and Zhejiang Province, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, China.
出版信息
Virol J. 2018 Jun 25;15(1):105. doi: 10.1186/s12985-018-1014-7.
BACKGROUND
Rice stripe virus (RSV) belongs to the genus Tenuivirus. It is transmitted by small brown planthoppers in a persistent and circulative-propagative manner and causes rice stripe disease (RSD). The NS3 protein of RSV, encoded by the viral strand of RNA3, is a viral suppressor of RNA silencing (VSR). NS3 plays a significant role in viral infection, and NS3-transgenic plants manifest resistance to the virus.
METHODS
The stability and availability of NS3 produced by transgenic Nicotiana benthamiana was investigated by northern blot analysis. The accumulation of virus was detected by western blot analysis. Transcriptome sequencing was used to identify differentially expressed genes (DEGs) in NS3-transgenic N. benthamiana.
RESULTS
When the host plants were inoculated with RSV, symptoms and viral accumulation in NS3-transgenic N. benthamiana were reduced compared with the wild type. Transcriptome analysis identified 2533 differentially expressed genes (DEGs) in the NS3-transgenic N. benthamiana, including 597 upregulated genes and 1936 downregulated genes. These DEGs were classified into three Gene Ontology (GO) categories and were associated with 43 GO terms. KEGG pathway analysis revealed that these DEGs were involved in pathways associated with ribosomes (ko03010), photosynthesis (ko00195), photosynthesis-antenna proteins (ko00196), and carbon metabolism (ko01200). More than 70 DEGs were in these four pathways. Twelve DEGs were selected for RT-qPCR verification and subsequent analysis. The results showed that NS3 induced host resistance by affecting host gene expression.
CONCLUSION
NS3, which plays dual roles in the process of infection, may act as a VSR during RSV infection, and enable viral resistance in transgenic host plants. NS3 from RSV affects the expression of genes associated with ribosomes, photosynthesis, and carbon metabolism in N. benthamiana. This study enhances our understanding of the interactions between VSRs and host plants.
背景
水稻条纹病毒（RSV）属于纤细病毒属。它通过褐飞虱以持久和循回半持久方式传播，引起水稻条纹病（RSD）。RSV 的 NS3 蛋白由 RNA3 病毒链编码，是一种病毒 RNA 沉默抑制子（VSR）。NS3 在病毒感染中起重要作用，而 NS3 转基因植物表现出对病毒的抗性。
方法
通过 northern blot 分析研究了 NS3 蛋白在转基因烟草原生质体中的稳定性和可用性。通过 western blot 分析检测病毒的积累。使用转录组测序鉴定 NS3 转基因烟草原生质体中的差异表达基因（DEGs）。
结果
当宿主植物被 RSV 接种时，与野生型相比，NS3 转基因烟草原生质体中的症状和病毒积累减少。转录组分析鉴定了 NS3 转基因烟草原生质体中的 2533 个差异表达基因（DEGs），包括 597 个上调基因和 1936 个下调基因。这些 DEGs 被分为三个基因本体（GO）类别，并与 43 个 GO 术语相关。KEGG 通路分析表明，这些 DEGs 参与与核糖体（ko03010）、光合作用（ko00195）、光合作用-天线蛋白（ko00196）和碳代谢（ko01200）相关的途径。这四个途径中超过 70 个 DEGs。选择 12 个 DEGs 进行 RT-qPCR 验证和后续分析。结果表明，NS3 通过影响宿主基因表达诱导宿主抗性。
结论
在感染过程中发挥双重作用的 NS3，在 RSV 感染过程中可能作为 VSR 发挥作用，并使转基因宿主植物具有抗病毒抗性。RSV 的 NS3 影响烟草原生质体中与核糖体、光合作用和碳代谢相关的基因表达。本研究增进了我们对 VSR 与宿主植物相互作用的理解。
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