Chamberland H, Charest P M, Ouellette G B, Pauzé F J
Histochem J. 1985 Mar;17(3):313-21. doi: 10.1007/BF01004593.
A cytochemical technique for the ultrastructural localization of chitin in tomato root cells infected by Fusarium oxysporum f. sp. radicis-lycopersici is reported. Chitinase was complexed to colloidal gold and thin sections were incubated with the enzyme-gold complex. This technique yielded a more uniform distribution of gold particles over the fungus wall, compared to that obtained with the lectin-gold technique. Both techniques revealed no labelling of the fungus cytoplasm, except for organelles resembling Woronin bodies. No significant labelling of either healthy or infected root cells was seen except for the secondary walls of vessels and, occasionally, that of adjoining parenchyma cells. The importance of this technique in studying the development of the pathogen within host cells is discussed.
报道了一种用于在被尖孢镰刀菌番茄根专化型感染的番茄根细胞中对几丁质进行超微结构定位的细胞化学技术。将几丁质酶与胶体金复合,并用酶-金复合物孵育薄切片。与凝集素-金技术相比,该技术在真菌壁上产生了更均匀的金颗粒分布。两种技术均未显示真菌细胞质有标记,除了类似沃罗宁体的细胞器。除了导管的次生壁以及偶尔相邻薄壁细胞的次生壁外,未观察到健康或受感染根细胞有明显标记。讨论了该技术在研究病原体在宿主细胞内发育中的重要性。