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通过 ESI-MS/MS 鉴定感染大豆根的大豆疫霉菌防御相关细胞壁蛋白。

Identification of defence-related cell wall proteins in Phytophthora sojae-infected soybean roots by ESI-MS/MS.

机构信息

Department für Biologie I der Universität, Botanik, Menzinger Str. 67, D-80638 München, Germany.

出版信息

Mol Plant Pathol. 2002 May 1;3(3):163-6. doi: 10.1046/j.1364-3703.2002.00109.x.

Abstract

Summary Root cell wall proteins involved in the incompatible interaction between soybean (Glycine max L.) and the phytopathogenic oomycete Phytophthora sojae were investigated by a proteomic approach using ESI-MS/MS analysis. Success of infection and subsequently induced defence reactions was proven by staining the cell wall localized defence-related 1,3-beta-glucan, callose, in semi-thin sections of the roots. Cell wall proteins of roots were extracted three weeks post-infection, separated by SDS-PAGE and eight selected protein bands were digested with trypsin. The resulting peptides were subjected to mass spectrometry-based sequencing. For four proteins, amino acid sequence information was obtained and used to identify the corresponding proteins by a homology search in databases. All four proteins (cationic peroxidase, peroxidase precursor, amine oxidase and lipoxygenase) were defence-related and their involvement in the oxidative burst is discussed. Three of these proteins have not been described for soybean before.

摘要

采用 ESI-MS/MS 分析的蛋白质组学方法研究了大豆(Glycine max L.)与植物病原卵菌大豆疫霉之间不亲和相互作用中涉及的根细胞壁蛋白。通过对根部半薄切片中细胞壁定位的防御相关 1,3-β-葡聚糖、几丁质的染色,证明了感染的成功和随后诱导的防御反应。感染后三周提取根部细胞壁蛋白,用 SDS-PAGE 分离,用胰蛋白酶消化 8 个选定的蛋白条带。所得肽段进行基于质谱的测序。对于四种蛋白质,获得了氨基酸序列信息,并通过数据库中的同源性搜索用于鉴定相应的蛋白质。这四种蛋白质(阳离子过氧化物酶、过氧化物酶前体、胺氧化酶和脂氧合酶)均与防御反应有关,并且讨论了它们在氧化爆发中的作用。其中三种蛋白质以前未在大豆中描述过。

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