马铃薯核糖体蛋白参与烟草脉带花叶病毒侵染的分子机制

植物病毒需要寄主蛋白的帮助才能进行复制和移动，进而危害作物安全生产。鉴定与病毒蛋白互作的植物蛋白，明确其在病毒侵染中的作用和机制，对设计新的作物病毒病防治策略有重要意义。烟草脉带花叶病毒（TVBMV）是马铃薯上的新发病毒，在生产中的危害逐年上升。申请者发现本氏烟核糖体蛋白L1可与TVBMV复制酶NIb互作，沉默核糖体蛋白L1编码基因能抑制TVBMV对本氏烟的侵染。本项目拟鉴定马铃薯与TVBMV NIb特异互作的核糖体蛋白种类、最小结构域和关键氨基酸，阐明马铃薯核糖体蛋白在TVBMV侵染中的作用和分子机制，利用CRISPR/Cas9技术对核糖体蛋白关键氨基酸位点进行编辑，分析编辑后马铃薯植株对TVBMV和相关病毒的抗性。预期结果将揭示马铃薯核糖体蛋白在TVBMV侵染中的作用和分子机制，发现新的抗病毒靶标，为抗TVBMV马铃薯新种质培育及相关病毒的可持续防治提供理论依据。

Plant viruses cause huge economic losses to crop production in the world. However, plant viruses must recruit host protein(s) to replicate, move, and establish systemic infection. Identifying the plant protein(s) interacting with viral proteins and elucidating their role in and mechanism of pathogenesis will be insightful to explore new strategies for plant viral disease control. Tobacco vein banding mosaic virus (TVBMV; Genus Potyvirus) is an emerging virus of potato and causes significant economic losses to potato production. In the previous study, we showed that the ribosomal protein L1 (RPL1) of Nicotiana benthamiana could interact with nuclear inclusion body b (NIb), the RNA-dependent RNA polymerase of TVBMV; silencing RPL1 gene inhibited the systemic infection of TVBMV in Nicotiana benthamiana plants. In this program, we plan to identify the species, minimum domain and critical amino acids of potato ribosomal protein(s) that specifically interact with TVBMV NIb using yeast two hybrid system (YTHS), bi-molecular fluorescence complementation (BiFC) and co-immunoprecipitation (Co-IP) assays, analyze the roles of potato ribosomal protein(s) in the pathogenesis of TVBMV by over-expressing and silencing the corresponding ribosomal protein gene, dissect the underlying molecular mechanisms, engineer potato plants by editing the ribosomal protein amino acids critical for the interaction with TVBMV NIb via CRISPR/Cas9, and analyze their resistance to TVBMV and related potyviruses. The expected results will elucidate the role of and mechanism underlying potato ribosomal protein(s) regulating infection of TVBMV and other potyviruses, reveal novel antiviral targets, and provide theoretical bases for the breeding of TVBMV- resistant potato plants. The expected results will also offer a scientific guide for the sustainable control of other potyviruses.