Perbal M C, Thomas C L, Maule A J
Department of Virus Research, John Innes Institute, John Innes Centre, Norwich, United Kingdom.
Virology. 1993 Jul;195(1):281-5. doi: 10.1006/viro.1993.1375.
Immunofluorescent staining of Chinese cabbage protoplasts infected with cauliflower mosaic virus (CaMV) showed different patterns of staining when antibodies raised against virus particles or antibodies specific for the CaMV movement protein (P1) were used. Anti-virion serum showed that particles accumulated as small discrete aggregates within the cytoplasm of infected protoplasts. In contrast, anti-P1 serum identified long thread-like structures extending from the protoplast surface. The same structures were seen when protoplasts were isolated from infected plants and held in culture. Electron microscopical examination after immunogold labeling and negative staining of protoplasts revealed some extracellular tubular structures which labeled with anti-P1 and which contained virus particles. Western blot analysis of isolated tubules showed them to be composed of the full-length P1 but not the smaller P1 products detected in infected tissues or protoplasts. The time course of P1 accumulation in infected protoplasts paralleled the accumulation of virus coat protein, as measured by immunofluorescence microscopy, and did not indicate that P1 was an early gene product as has been suggested for other virus movement proteins.
用针对花椰菜花叶病毒(CaMV)病毒粒子产生的抗体或针对CaMV运动蛋白(P1)的特异性抗体对感染CaMV的大白菜原生质体进行免疫荧光染色时,显示出不同的染色模式。抗病毒粒子血清显示,病毒粒子在受感染原生质体的细胞质中聚集成小的离散聚集体。相比之下,抗P1血清鉴定出从原生质体表面延伸出的长丝状结构。当从受感染植物中分离原生质体并进行培养时,也能看到同样的结构。对原生质体进行免疫金标记和负染色后的电子显微镜检查发现了一些细胞外管状结构,这些结构用抗P1抗体标记并含有病毒粒子。对分离出的管状结构进行蛋白质免疫印迹分析表明,它们由全长P1组成,而不是在受感染组织或原生质体中检测到的较小的P1产物。通过免疫荧光显微镜测量,受感染原生质体中P1积累的时间进程与病毒外壳蛋白的积累平行,并且没有表明P1是如其他病毒运动蛋白所暗示的早期基因产物。