酵母双杂交筛选与小麦黄花叶病毒P2互作的寄主因子  

Screening of host factors interacting with wheat yellow mosaic virus P2 by yeast two-hybrid system

作　　者：韩晓蕾 高仕祺 张帆[3] 羊健 刘芃 姜鸿明[1] 李林志[1] HAN Xiaolei;GAO Shiqi;ZHANG Fan;YANG Jian;LIU Peng;JIANG Hongming;LI Linzhi(Yantai Academy of Agricultural Sciences, Yantai 265500, China;College of Life Sciences, Yantai University, Yantai 265500, China;Ningbo University/State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Ningbo 315000, China)

机构地区：[1]烟台市农业科学研究院,山东烟台265500 [2]烟台大学生命科学学院,山东烟台265500 [3]宁波大学/农产品质量安全危害因子与风险防控国家重点实验室,浙江宁波315000

出　　处：《浙江农业学报》2021年第3期497-505,共9页Acta Agriculturae Zhejiangensis

基　　金：山东省农业良种工程(2019LZGC001);山东省重大科技创新工程(2018YFJH0602);国家重点研发计划(2017YFD0100600);农业部财政部项目(CARS-3-2-23);丘陵旱地小麦-玉米抗旱减灾技术集成与示范。

摘　　要：小麦黄花叶病毒(wheat yellow mosaic virus,WYMV)隶属马铃薯Y病毒科(Potyviridae),大麦黄花叶病毒属(Bymovirus)。该病毒基因组是由两条正义单链RNA1和RNA2构成,共编码10个蛋白,其编码的P2在病毒的复制过程中发挥重要功能。文章构建了小麦cDNA酵母文库,通过共转试验共筛选获得若干个候选互作寄主因子,包括水杨酸信号通路因子、E3泛素化连接酶、防御反应、氧化还原酶活性、光系统II组件等18种蛋白,研究P2与寄主因子的互作。进一步试验证实,其中的TaTIFY 10A、Ta14-3-3与WYMV-P2存在互作关系。结果表明,WYMV-P2与小麦的E3泛素化类转录因子、水杨酸信号转录因子、植物光系统的稳定组件及叶绿体的形成关键因子等可能存在互作关系,P2可能参与了寄主多种信号途径,为明确WYMV与寄主的互作机制提供了研究基础。Wheat yellow mosaic virus(WYMV)is classified in the genus Bymovirus,family Potyviridae.The genome is composed of two sense RNA single-stranded RNA1 and RNA2 and encodes 10 proteins,among which P2 plays an important role in virus replication.In order to identify the targets of P2 in host,co-transformation of cDNA library vector and pGBKT7-P2 was constructed,eighteen candidate host targets of P2 were screened out,which were divided into several types based on their functions,including salicylic acid signal pathway factor,E3 ubiquitin ligase,defense response,oxidoreductase activity and photosystem II assembly.Additionally,the interaction of P2 and TaTIFY or Ta14-3-3 was further verified by Y2H system.Based on the results,it could be speculated that P2 may interact with E3 ubiquitin transcription factor,salicylic acid signal transcription factor,or the factors associated with plant photosystem and chloroplasts formation.In conclusion,P2 may participate in multiple signaling pathways during viral infection,and provide an insight into molecular interaction mechanism between WYMV and the host.

关 键 词：小麦黄花叶病毒 小麦 P2 CDNA文库 酵母双杂交 

分 类 号：S435.12[农业科学—农业昆虫与害虫防治]