Ruiz-Herrera J, Leon C G, Carabez-Trejo A, Reyes-Salinas E
Department of Genética y Biología Molecular, Unidad México, D.F., México.
Fungal Genet Biol. 1996 Jun;20(2):133-42. doi: 10.1006/fgbi.1996.0028.
Isolated or in vivo cell walls from the yeast and mycelial forms of haploid Ustilago maydis were not stained by the normal osmium procedure for electron microscopy. KMnO4 stained mycelial walls, revealing a layered structure with a loose electron-dense layer at the cell surface, but stained only the outer surface layer of yeast walls. Walls were purified from extracts obtained by ballistic and ultrasonic disruption. Chemical analysis showed that composition of yeast and mycelial walls was similar. Yeast walls contained higher amounts of neutral sugars and protein, whereas mycelial walls contained more chitin and phosphate. No chitosan or uronic acids were detected. Higher proportions of xylose and mannose were present in yeast walls, whereas the amounts of glucose and galactose were higher in mycelial walls. Fucose, arabinose, and ribose were detected in yeast walls only. Electrophoretic patterns of proteins extracted with SDS, beta 1, 3-glucanase, or chitinase were similar in walls of both morphologies, although some differential bands were identified. Most antigenic proteins appeared in the covalently bound fraction of the wall. Some were common to both morphologies, but others were stage specific.
单倍体玉米黑粉菌酵母型和菌丝型的离体或体内细胞壁,用常规的锇酸电子显微镜染色法无法染色。高锰酸钾可对菌丝细胞壁染色,显示出一种分层结构,在细胞表面有一层松散的电子致密层,但只能对酵母细胞壁的外层进行染色。细胞壁是从通过高速撞击和超声破碎获得的提取物中纯化得到的。化学分析表明,酵母细胞壁和菌丝细胞壁的组成相似。酵母细胞壁含有较高含量的中性糖和蛋白质,而菌丝细胞壁含有更多的几丁质和磷酸盐。未检测到壳聚糖或糖醛酸。酵母细胞壁中木糖和甘露糖的比例较高,而菌丝细胞壁中葡萄糖和半乳糖的含量较高。岩藻糖、阿拉伯糖和核糖仅在酵母细胞壁中检测到。用SDS、β-1,3-葡聚糖酶或几丁质酶提取的蛋白质的电泳图谱在两种形态的细胞壁中相似,尽管鉴定出了一些差异条带。大多数抗原蛋白出现在细胞壁的共价结合部分。有些是两种形态共有的,但其他的则具有阶段特异性。