Grenier J, Potvin C, Asselin A
Département de Phytologie, Faculté des Sciences de l'Agriculture et de l'Alimentation, Université Laval, Québec, Canada.
Plant Physiol. 1993 Dec;103(4):1277-83. doi: 10.1104/pp.103.4.1277.
Proteins from intercellular fluid extracts of chemically stressed barley (Hordeum vulgare L.) leaves were separated by native polyacrylamide gel electrophoresis at alkaline or acid pH. Polyacrylamide gels contained Saccharomyces cerevisiae (bakers' yeast) or Schizosaccharomyces pombe (fission yeast) crude cell walls for assaying yeast wall lysis. In parallel, gels were overlaid with a suspension of yeasts for assaying growth inhibition by pathogenesis-related proteins. The same assays were also performed with proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under nonreducing conditions. In alkaline native polyacrylamide gels, only one band corresponding to yeast cell wall lytic activity was found to be inhibitory to bakers' yeast growth, whereas in acidic native polyacrylamide gels one band inhibited the growth of both yeasts. Under denaturing nonreducing conditions, one band of 19 kD inhibited the growth of both fungi. The 19-kD band corresponded to a basic protein after two-dimensional gel analysis. The 19-kD protein with yeast cell wall lytic activity and inhibitory to both yeasts was found to be different from previously reported barley chitosanases that were lytic to fungal spores. It could be different from other previously reported lytic antifungal activities related to pathogenesis-related proteins.
通过在碱性或酸性pH条件下进行非变性聚丙烯酰胺凝胶电泳，分离化学胁迫的大麦（Hordeum vulgare L.）叶片细胞间液提取物中的蛋白质。聚丙烯酰胺凝胶含有酿酒酵母（面包酵母）或粟酒裂殖酵母（裂殖酵母）的粗细胞壁，用于检测酵母细胞壁裂解。同时，凝胶用酵母悬液覆盖，用于检测病程相关蛋白对酵母生长的抑制作用。对在非还原条件下通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离的蛋白质也进行了相同的检测。在碱性非变性聚丙烯酰胺凝胶中，仅发现一条对应酵母细胞壁裂解活性的条带对面包酵母生长有抑制作用，而在酸性非变性聚丙烯酰胺凝胶中，一条条带抑制两种酵母的生长。在变性非还原条件下，一条19 kD的条带抑制两种真菌的生长。二维凝胶分析后，19 kD的条带对应一种碱性蛋白。发现具有酵母细胞壁裂解活性且对两种酵母都有抑制作用的19 kD蛋白与先前报道的对真菌孢子有裂解作用的大麦壳聚糖酶不同。它可能与先前报道的其他与病程相关蛋白有关的裂解抗真菌活性不同。
