植物病毒粒体在寄主细胞中的原位聚集特征  

Plant virions in situ aggregated characterization in the host cells

作　　者：张仲凯[1] 郑宽瑜[1] 王田田 吴少政 裴卫华[1] ZHANG Zhong-kai;ZHENG Kuan-yu;WANG Tian-tian;WU Shao-zheng;PEI Wei-hua(Institute of Biotechnology and Germplasm Resources,Yunnan Academy of Agricultural Sciences,Key Laboratory of Agricultural Biotechnology of Yunnan Province,Kunming Yunnan 650223,China)

机构地区：[1]云南省农业科学院生物技术与种质资源研究所云南省农业生物技术重点实验室

出　　处：《电子显微学报》2019年第5期531-541,共11页Journal of Chinese Electron Microscopy Society

基　　金：国家自然科学基金-云南联合基金重点资助项目(No.U1802235);云南省云岭学者培养计划项目

摘　　要：病毒粒体的原位聚集特征是快速诊断植物病毒及研究其系统侵染的基础,但国内外对此研究和报道较少。本研究采用马铃薯Y病毒属病毒、烟草花叶病毒、番茄斑萎病毒属病毒、水稻矮缩病毒、黄瓜花叶病毒等不同形态的植物病毒自然感染或人工接种的植株样品,比较了研磨过滤、蔗糖密度梯度离心、切片浸渍法、超薄切片与高压冷冻等不同电镜制样方法观察效果。结果表明,以本实验室发明的切片浸渍(原生质体裂解)法负染色制样与超薄切片制样透射电镜观察,可以得到线状、杆状和被膜球状植物病毒粒体在寄主细胞中的典型原位聚集特征,借此可区分不同属或种,对植物病毒的快速诊断鉴定具有重要参考价值。在此基础上,对应用植物病毒粒体原位聚集特征深入研究病毒复制、装配与运动的机制进行探讨,为植物病毒的系统侵染机理研究提供依据。Virions in situ aggregated characterization is the basis for the rapid diagnosis of plant virus and the study of their systemic infection,but its researches is limited.In this study,different plant viral samples with various morphology,including potyviruses,Tobacco mosaic virus,tospoviruses,Rice dwarf virus,Cucumber mosaic virus,were collected from the field and mechanical inoculated in new plant.Then observation results of electron microscopy with different preparation methods,such as grinding filter,sucrose density gradient centrifugation,slice impregnated negative staining,ultrathin section,and high pressure freeze-electron microscopy,were compared.The results indicated that the typical in situ aggregated characterization of the filamentousviruses,baculoviruses and sphericalviruses in host cells could be obtained by slice impregnated method(protoplast cracking)developed by our laboratory combined with ultrathin section.These virions in situ aggregated characterization could be helpful to distinguish plant virus from genus or species and had important reference value for rapid diagnosis and identification of plant viruses.On the above results,it would be helpful to study virus replication,assembly,and movement mechanism according to virions in situ aggregated characterization,so as to provide a basis for the study of systemic infection mechanism of plant viruses.

关 键 词：植物病毒粒体 样品制备 透射电镜 原位聚集 原生质体裂解 

分 类 号：S432.41[农业科学—植物病理学]