应用免疫胶体金技术定位感病大麦细胞中的大麦和性花叶病毒

本文报道免疫胶体金标记技术的建立,并用此技术定位大麦叶和根组织超薄切片中大麦和性花叶病毒(BaMMV)。在感染病毒的大麦叶和根细胞中,病毒束、游离病毒颗粒和病毒外壳蛋白多分布于细胞质丰富的细胞中,且以液泡和叶绿体(仅叶组织)周围较多。在细胞器已解体的病根表皮细胞中,有时也可检测到大量游离病毒粒子。少数风轮体或板状集结体上也存在病毒或病毒外壳蛋白。细胞核、叶绿体、线粒体、细胞膜以及其他细胞器上都未见有特异性金颗粒标记。

Localization of Barley Mild Mosaic Virus in Infected Barley Cells by Immuno-Colloidal Gold Labelling

An immuno-colloidal gold labelling procedure was developed to localize barley mild mosaic virus(BaMMV) in thin sections of barley leaf and root tissues embedded in L. R. White resin, The optimal procedure for specific labelling of virus with a minimum of background labelling and economical use of reagents was to treat sections with buffer containing bovine serum albumen to block non-specific binding sites, treat with BaMMV-IgG(1/1000; 15 rain at room temperature), wash and label with goat anti-rabbit IgG-gold conjugate(1/80; 5 min at room temperature). After a further washing, sections were stained with uranyl acetate and lead citrate. In both leaves and roots, virus bundles, free virus particles and coat protein were localized in the cytoplasm, usually near to vacuole membranes and, in leaf tissue, chloroplasts, Some virus bundles and coat protein were also bound onto pinwheels or laminated aggregates but they were rarely seen on membranous bodies, No specific labelling was seen on nuclei, chloroplasts, mitochondria, cell walls or other organelles, In roots, virus was concentrated near the vascular bundles and was also sometimes seen in degenerating epidermal cells.