CMV-IB侵染诱导烟草内质网应激  

Cucumber Mosaic Virus-IB Induces Endoplasmic Reticular Stress in Nicotiana

作　　者：董文凤 李方方[1] 何青云 肖志新 孙航军 王升平[1] 杨金广[1] 王凤龙[1] 申莉莉[1] DONG Wenfeng;LI Fangfang;HE Qingyun;XIAO Zhixin;SUN Hangjun;WANG Shengping;YANG Jinguang;WANG Fenglong;SHEN Lili(Tobacco Research Institute, Chinese Academy of Agricultural Science, Qingdao 266101, China;Baoshan Tobacco (Group) Co., Ltd., Baoshan, Yunnan 678000, China)

机构地区：[1]中国农业科学院烟草研究所,青岛266101 [2]云南省烟草公司保山市公司,云南保山678000

出　　处：《中国烟草科学》2017年第6期1-11,共11页Chinese Tobacco Science

基　　金：山东省自然科学基金"PVY HC-Pro与番茄CAT互作在PVY致病中的作用分析"(ZR2014CQ025);云南省烟草公司科技项目"保山烟区主要病虫害成灾预警与绿色防控技术研究与应用"(2016YN12);红云红河烟草(集团)有限责任公司科技项目"提高保山基地烟叶品质的关键技术研究及应用"(HYHH2016YL02)

摘　　要：病毒复制产生的大量病毒蛋白阻碍寄主细胞转运功能,未/错折叠蛋白在内质网腔蓄积导致内质网应激(ER stress)。为明确利用液泡膜复制的黄瓜花叶病毒(CMV)是否诱导寄主ER stress,在CMV-IB侵染三生烟和本氏烟体系中,通过qRT-PCR检测ER stress相关基因的转录水平,农杆菌介导ER marker(m Cherry-HDEL)瞬时转染后激光共聚焦显微观察ER形态,电镜观察细胞病理。结果显示,较PBS对照,接种CMV后12 h,ER stress相关基因的转录水平显著上调1.23~7.37倍;5~10 d ER膜微丝发生明显的重排和增生;病变细胞叶绿体降解、线粒体肿胀,且沿液泡膜的微自噬和细胞质中的巨自噬结构增多。表明CMV侵染通过激活ER stress基因和改变ER膜形态诱导寄主ER stress,寄主细胞会以细胞器降解、自噬乃至凋亡的方式抵御病毒扩展。The cellular translation machinery is hijacked by large amounts of viral proteins upon virus infection.The unfolded proteins accumulated in the endoplasmic reticulum(ER)results in ER stress.ER stress is implicated in some plant virus infections that replicated in the ER.To investigate if Cucumber mosaic virus(CMV)that replicated in the tonoplast induces ER stress,ER stress gene expression by qRT-PCR,ER morphology by transient expression of mCherry-HDEL and confocal microscopy,and ultrastructure of cytopathology by electron microscope in CMV-IB inoculated Nicotiana leaves were compared with PBS buffer inoculated control.The results showed that the ER stress related genes were upregulated significantly at12hours post inoculation,with1.23-7.37folds higher levels in CMV inoculated leaves than in PBS controls(p<0.05).ER morphology was hyperplasia and significant rearrangement at5-10days post CMV inoculation.Ultrastructure of CMV-diseased cells showed chloroplast degradation,mitochondria swelling,invagination of tonoplast into two double membranous complex structures of microautophagy,and macroautophagic structures in the cytoplasm.This indicated that CMV infection induces ER stress including activating ER stress genes and causing ER proliferation.Host may activate organelles degradation,autophagy even cell death to suppress viral infection.

关 键 词：黄瓜花叶病毒 内质网应激 细胞病理 自噬 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]