叶锈菌侵染的小麦细胞间隙液中激发子的分离纯化

Purification of an Elicitor from Intercellular Washing Fluids of Rust-Infected Wheat Leaves

【目的】在已经证明小麦与叶锈菌互作的小麦细胞间隙液(IWF)中存在具有蛋白质性质的激发子活性物质的基础上,以期通过试验得到激发子的纯品制剂,为进一步研究激发子的组成和结构、激发子受体以及寄主产生过敏性反应的信号转导机制提供基础条件。【方法】提取叶锈菌小种165侵染的小麦品种洛夫林10细胞间隙液,通过盐析、凝胶过滤柱层析和离子交换柱层析等分离纯化技术,对IWF中具有激发子活性的蛋白质组分进行分离纯化。【结果】分离出能诱导洛夫林10健康叶片PAL、PO活性增高并诱发洛夫林10悬浮细胞程序性死亡的激发子活性组分。该组分经SDS-PAGE电泳呈现单一条带,分子量约为32kD。【结论】经硫酸铵分段盐析、凝胶过滤柱层析和离子交换柱层析等方法,从感染叶锈菌小种165的洛夫林小麦叶片细胞间隙液中分离出具有激发子活性的蛋白质组分。

[ Objective ] In the prevenient studies, there was an evidence that a proteinaceous elicitor in the intercellular washing fluids （1WF） extracted from wheat leaves inoculated with leaf rust fungus （Puccinia triticina）. In order to obtain the purified elicitor, a study was made and the results are expected to provide a molecular basis for further exploring the elicitor＇s characteristics, receptor and parasitifer＇s hypersensitive response signal transduction system. [ Method ] IWF was extracted from wheat leaves （cultivar lovrin 10） inoculated with leaf rust race 165 and then purified through the procedures of salting out, gel-filtration chromatography and DEAE anion exchange chromatography. [Result] The activity of defense-related enzymes, such as PAL and PO, was increased in healthy wheat leaves （lovrin 10） treated with the elicitor. The elicitor also induced PCD arising in suspension cell of lovrin 10. [Conclusion] The purified elicitor＇s molecular weight is about 32 kDa in SDS -PAGE.