Extracellular beta-1,3-Glucanases in Stem Rust-Affected and Abiotically Stressed Wheat Leaves : Immunocytochemical Localization of the Enzyme and Detection of Multiple Forms in Gels by Activity Staining with Dye-Labeled Laminarin.
Extracellular beta-1,3-Glucanases in Stem Rust-Affected and Abiotically Stressed Wheat Leaves : Immunocytochemical Localization of the Enzyme and Detection of Multiple Forms in Gels by Activity Staining with Dye-Labeled Laminarin.
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DOI：
10.1104/pp.94.3.1376
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发表时间：
1990-11
期刊：
影响因子：
7.4
通讯作者：
J. Sock;R. Rohringer;Z. Kang
中科院分区：
文献类型：
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作者：
J. Sock;R. Rohringer;Z. Kang
小麦（Triticum aestivum）叶子感染小麦茎锈菌（Puccinia graminis f.sp. tritici）后4天，其细胞间冲洗液（IWF）中的内切-β-1,3-葡聚糖酶活性增加了10倍，而外切-β-1,3-葡聚糖酶活性保持在较低水平不变。热和乙烯胁迫没有影响，而汞处理导致内切 β-1,3-葡聚糖酶活性增加 2 倍。通过使用昆布多糖-雷马素亮蓝作为覆盖凝胶中的底物进行活性染色的新方法，在未受胁迫的叶子的 IWF 中检测到了 18 种内切 β-1,3-葡聚糖酶，在受茎锈病感染的叶子的 IWF 中检测到了多达 24 种形式。锈病感染后 β-1,3-葡聚糖酶活性和 β-1,3-葡聚糖酶数量的增加大部分是由于对植物的非特异性、与胁迫相关的影响，但该酶的两种主要形式可能源自真菌。用抗大麦-β-1,3-葡聚糖酶抗体对β-1,3-葡聚糖酶进行细胞化学定位。通过预嵌入标记，该酶在宿主和真菌细胞壁的外部得到证实。嵌入后标记将酶定位在宿主质膜中和邻近质膜的宿主细胞壁区域中、细胞间菌丝细胞和吸器的整个壁、真菌细胞质中以及吸器外基质中。来自小麦的 β-1,3-葡聚糖酶和锈菌发芽的黑孢子与抗大麦-β-1,3-葡聚糖酶抗体的交叉反应性在斑点印迹测定和蛋白质印迹中得到证实。
Endo-beta-1,3-glucanase activity in intercellular washing fluid (IWF) from leaves of wheat (Triticum aestivum) increased 10-fold 4 days after leaves were infected with the wheat stem rust fungus (Puccinia graminis f.sp. tritici), while exo-beta-1,3-glucanase activity remained unchanged at a low level. Heat and ethylene stress had no effect, whereas mercury treatment resulted in a 2-fold increase in endo-beta-1,3-glucanase activity. With a new method of activity staining using laminarin-Remazol brilliant blue as substrate in overlay gels, 18 electrophoretic forms of endo-beta-1,3-glucanase were detected in IWF from unstressed leaves and up to 24 forms in IWF from stem rust-infected leaves. Most of the increase in beta-1,3-glucanase activity and in the number of beta-1,3-glucanases after rust infection was due to a nonspecific, stress-related effect on the plant, but two major forms of the enzyme probably originated from the fungus. beta-1,3-Glucanase was localized cytochemically with anti-barley-beta-1,3-glucanase antibodies. With preembedding labeling, the enzyme was demonstrated on the outside of host and fungal cell walls. Postembedding labeling localized the enzyme in the host plasmalemma and in the domain of host cell walls adjoining the plasmalemma, throughout walls of intercellular hyphal cells and haustoria, in the fungal cytoplasm, and in the extrahaustorial matrix. Cross-reactivity of beta-1,3-glucanases from wheat and germinated uredospores of the rust fungus with the anti-barley-beta-1,3-glucanase antibodies was confirmed in dot blot assays and on Western blots.