植物病毒在世界范围内对农业生产造成严重危害。BYL in vitro病毒翻译复制体系（BYL体系）是脱去液泡的BY-2烟草原生质体裂解液，适用于植物RNA病毒翻译、复制机制以及相关宿主蛋白质的研究。嘧肽霉素是由项目组所在沈阳农业大学自主开发，可有效抑制烟草花叶病毒（TMV）所引起的病害的生物药剂。本项目通过米台霉素和宁南霉素两种抗病毒生物药剂和TMV来详细研究生物药剂的抗病毒分子机理。本研究拟采用BY-2原生质体法研究抗病毒生物药剂对TMV在单细胞内的作用；拟采用BYL 体系明确生物药剂对TMV的翻译以及RNA负义链合成的影响；拟基于BYL体系下采用Strepto-Tag法和免疫共沉淀法明确上述药剂对与TMV相互作的致病相关宿主蛋白质的影响，并通过液相色谱串联质谱法对目标宿主蛋白进行鉴定，进而明确生物药剂对病毒相关宿主蛋白质的影响。本研究将为更有效专抗病毒药剂的开发提供理论依据和重要参考。

Plant viruses cause great losses to agriculture worldwide every year . BYL (evacuolated tobacco BY-2 protoplast Lysate) in vitro virus translation/replication system is widely applied to study the mechanisms of plant viral protein translation, replication and the host proteins involved in pathogenicity. Cytosinpeptidemycin is a biological antiviral agent developed by our research group of ShenYang agricultural University, which has good inhibitory effect on Tobacco mosaic virus (TMV). In this project, two biological antiviral agents of Ningnanmycin and Cytosinpeptidemycin together with TMV are used to further study their molecular antiviral mechanisms. In this study, Effect of antiviral agents on TMV accumulation in the single cell will be studied using BY-2 tobacco protoplast experiment. BYL in vitro system will be used to study the effect of the biological antiviral agents on TMV translation and negative RNA synthesis in separate experiments. Meanwhile, BYL system will also be used to study the effect of the biological antiviral agents on host proteins that is interacted with TMV RNA and P126 using Strepto-Tag and Co-IP method. The related host proteins will be identified through LC-MS/MS method. The studies of this project could provide important theoretical basis for the development of more specific and effective antiviral agents