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当前位置: 首页 > 文章 > 侵染朱槿的木尔坦棉花曲叶病毒V2蛋白的亚细胞定位特征分析 园艺学报 2018 (12) 2417-2426
Position: Home > Articles > Characterization and Subcellular Localization of V2 Protein Encoded by Cotton leaf curl Multan virus Infecting Hibiscus rosa-sinensis Acta Horticulturae Sinica 2018 (12) 2417-2426

侵染朱槿的木尔坦棉花曲叶病毒V2蛋白的亚细胞定位特征分析

作  者：
陈思敏;季英华;吴根土;赵文浩;吴淑华;肖留斌;孙现超;周益军;青玲
单  位：
西南大学植物保护学院植物病害生物学重庆市高校级重点实验室;江苏省农业科学院植物保护研究所江苏省食品质量安全重点实验室—省部共建国家重点实验室培养基地
关键词：
朱槿;木尔坦棉花曲叶病毒;V2蛋白;亚细胞定位
摘  要：
木尔坦棉花曲叶病毒(Cotton leaf curl Multan virus,CLCuMuV)2006年在中国首次报道,并呈逐年扩散蔓延,给园艺和经济作物造成了严重损失。选择2012年采自江苏南京的CLCuMuV朱槿(Hibiscus rosa-sinensis)分离物作为研究对象,对其V2蛋白基因进行了扩增及克隆,并构建了V2-YFP融合表达载体,利用农杆菌浸润法接种本氏烟(Nicotiana benthamiana)。激光共聚焦显微镜观察结果显示浸润处理后本氏烟叶片细胞的细胞质和细胞核周都有较强的绿色荧光信号,同时细胞质中还分布有点状的荧光信号。反转录PCR(RT-PCR)及蛋白质免疫印迹(Westernblot)结果显示带荧光标签的V2在本氏烟叶片中转录和表达正常。这些结果说明CLCuMuV编码的V2蛋白主要分布在细胞质和细胞核周,同时在细胞质中还会形成小聚合体。
译  名：
Characterization and Subcellular Localization of V2 Protein Encoded by Cotton leaf curl Multan virus Infecting Hibiscus rosa-sinensis
作  者：
CHEN Simin;JI Yinghua;WU Gentu;ZHAO Wenhao;WU Shuhua;XIAO Liubin;SUN Xianchao;ZHOU Yijun;QING Ling;College of Plant Protection,Southwest University/Chongqing Key Laboratory of Plant Disease Biology;Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Key Lab of Food Quality and Safety of Jiangsu Province-State Key Laboratory Breeding Base;
单  位：
CHEN Simin%JI Yinghua%WU Gentu%ZHAO Wenhao%WU Shuhua%XIAO Liubin%SUN Xianchao%ZHOU Yijun%QING Ling%College of Plant Protection,Southwest University/Chongqing Key Laboratory of Plant Disease Biology%Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Key Lab of Food Quality and Safety of Jiangsu Province-State Key Laboratory Breeding Base
关键词：
Hibiscus rosa-sinensis;;Cotton leaf curl Multan virus;;V2 protein;;subcellular localization
摘  要：
Cotton leaf curl Multan virus(CLCuMuV),an important whitefly-transmitted geminivirus,was first reported in China in 2006 and the epidemics of the virus caused devastating losses to local horticultural and economical crops in recent years. As V2 was reported essential for virus infection for many begomoviruses,to clarify the subcellular localization of V2 encoded by CLCuMuV,the V2 gene of a Jiangsu isolate of CLCuMuV from the Hibiscus rosa-sinensis,was amplified and cloned. Subsequently V2 was cloned into a 35 S:YFP vector to obtain YFP tagged V2(V2-YFP)and transiently expressed in the Nicotiana benthamiana leaves by Agrobacterium infiltration. The subcellular localization of V2 was observed by confocal laser scanning microscope and the result showed that there was relatively strong GFP fluorescence signal around the nucleus and at the cell periphery(cytoplasm),and some punctate fluorescent bodies were also observed in the cytoplasm. The expression of V2 protein(fused with YFP)in the infiltrated N. benthamiana leaves was confirmed by RT-PCR and Western blotting. These results demonstrated that the V2 protein encoded by CLCuMuV was localized around the nucleus and at the cell periphery(cytoplasm),and V2 protein could form aggregates in the cytoplasm as well.

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