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NS3 Protein from Rice stripe virus affects the expression of endogenous genes in Nicotiana benthamiana

机译:水稻条纹病毒的NS3蛋白影响本氏烟草内源基因的表达

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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. 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. 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. 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.
机译:水稻条纹病毒(RSV)属于Tenuivirus属。它由小的褐飞虱以持续和循环繁殖的方式传播,并引起水稻条纹病(RSD)。 RSV的NS3蛋白由RNA3的病毒链编码,是RNA沉默(VSR)的病毒抑制剂。 NS3在病毒感染中起重要作用,而NS3转基因植物表现出对该病毒的抗性。通过Northern印迹分析研究了转基因烟草本氏烟草产生的NS3的稳定性和可用性。通过蛋白质印迹分析检测病毒的积累。转录组测序用于鉴定NS3转基因本氏烟草中的差异表达基因(DEG)。当宿主植物接种RSV时,与野生型相比,NS3转基因本氏烟草中的症状和病毒积累减少了。转录组分析确定了NS3转基因本氏烟草中的2533个差异表达基因(DEG),包括597个上调基因和1936个下调基因。这些DEG分为三个基因本体论(GO)类别,并与43个GO术语相关联。 KEGG通路分析表明,这些DEG参与了与核糖体(ko03010),光合作用(ko00195),光合作用-天线蛋白(ko00196)和碳代谢(ko01200)相关的通路。这四个途径中有70多个DEG。选择十二个DEG用于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。在感染过程中起双重作用的NS3可能在RSV感染期间充当VSR,并在转基因宿主植物中实现病毒抗性。来自RSV的NS3影响本氏烟草中核糖体,光合作用和碳代谢相关基因的表达。这项研究增强了我们对VSR与寄主植物之间相互作用的理解。

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