TMV侵染烟草诱导寄主产生细胞自噬  

Tobacco mosaic virus infection on tobacco plants induces autophagy

作　　者：刘伟[1] 李方方[1] 孙航军 王耀锋 余广宏 王凤龙[1] 钱玉梅[1] 杨金广[1] 

机构地区：[1]中国农业科学院烟草研究所/烟草行业病虫害监测与综合治理重点开放实验室,青岛266101 [2]甘肃省庆阳市烟草公司,庆阳745000 [3]云南烟草保山香料烟有限责任公司,保山678000

出　　处：《植物病理学报》2016年第6期759-766,共8页Acta Phytopathologica Sinica

基　　金：烟草行业烟草病虫害监测与综合治理重点实验室专项经费

摘　　要：细胞自噬(autophagy)—自体吞噬,是真核生物中一种高度保守并普遍存在的物质循环利用过程。为明确烟草花叶病毒(Tobacco mosaic virus,TMV)侵染烟草是否引起细胞自噬,本研究利用TMV-U1和普通烟(Nicotiana tabacum cv.Blight yellow)为主要研究材料,利用透射电子显微镜观察、单丹磺酰尸胺(M DC)荧光染色技术、自噬标记分子ATG8f-ECFP荧光观察和荧光定量RT-PCR(qRT-PCR)分别检测TMV侵染烟草后的烟草细胞内自噬结构的形成、酸性细胞器的存在和细胞自噬相关基因(Autophagy related gene,ATG)ATG3、ATG4、ATG5、ATG6、ATG7、ATG8a、ATG18a的表达特征。结果表明,TM V侵染72 h后,在液泡边际和液泡内发现大量疑似自噬体结构。利用M DC荧光染色和ATG8f-ECFP瞬时表达技术,通过激光共聚焦显微镜观察,在TMV侵染72 h后发现点状荧光信号。TMV侵染48 h后,除ATG5外,烟草ATG3、ATG4、ATG6、ATG7、ATG8a、ATG18a表达均上调并在72 h达到顶峰。表明TM V侵染后感病烟草均可检测到细胞自噬现象,推测TMV侵染普通烟可诱导寄主产生细胞自噬反应。Autophagy （or self eating） is a highly conserved process by which eukaryotic cells recycle intracel- lular components. In this study, TMV-U1 and its susceptible host Nicotiana tabacum cv. Blight yellow, were used to elucidate whether Tobacco mosaic virus infection could induce autophagy in tobacco plants. Transmission electron microscope （TEM）, monodansylcadaverin （MDC） staining, confocal microscopy analysis of enhanced cyan fluorescent protein-tagged tobacco ATGSf （ECFP-ATGSf） and quantitative real-time PCR were used to in- vestigate the formation of autophagic structure, acidic compartments, and the transcript levels of autophagy rela- ted genes including ATG3, ATG4, ATG5, ATG6, ATG7, ATGSa, and ATG18a in TMV-infected tobacco plants, respectively. A number of autolysosome-like structures were observed in or near the periphery of the vac- uole at 72 hours post inoculation （hpi） under TEM. Strong punctate MDC-stained autolysosomes in the cells were obearse by comfacal cept ATG5, were increased at 48 hpi, and peaked 72 hpi. These results suggest that TMV infection induces auto- phagy in the susceptible tobacco plants.

关 键 词：烟草花叶病毒 感病寄主 细胞自噬 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]