Department of Plant Biochemistry, Dahlem Centre of Plant Sciences, Freie Universität Berlin, Königin-Luise-Str. 12-16, 14195, Berlin, Germany,
Plant Cell Rep. 2014 Nov;33(11):1865-79. doi: 10.1007/s00299-014-1663-y. Epub 2014 Jul 24.
Our studies indicate a potential important elicitor candidate which can aid in the fight against a worldwide disease, rice blast. In this study, we report the purification, identification, characterization, and gene cloning of a novel hypersensitive response-inducing protein elicitor (MoHrip2) secreted from an important pathogenic fungus, Magnaporthe oryzae. The protein fraction was isolated from the culture filtrate of M. oryzae and identified by de novo sequencing. The elicitor-encoding gene mohrip2 was cloned following sequence comparison and PCR amplification. This 459-bp gene encodes a 152-residue polypeptide that contains an 18-residue signal peptide and exhibits a pI of 4.72 and an apparent molecular mass of 16 kDa. The hypothetical protein, MoHrip2, was expressed in Escherichia coli, and both the recombinant and the endogenous protein caused necrotic lesions in tobacco leaves. In addition to phenolic compound deposition and alkalization of the extracellular medium, MoHrip2 also induced hydrogen peroxide production and nitric oxide accumulation in tobacco cells. Moreover, rice seedlings treated with MoHrip2 exhibited pronounced resistance to M. oryzae compared with control seedlings.
我们的研究表明,一种潜在的重要激发子候选物可以帮助对抗世界性疾病——稻瘟病。在这项研究中,我们报告了一种从重要病原菌稻瘟病菌中分泌的新型过敏反应诱导蛋白激发子(MoHrip2)的纯化、鉴定、特性和基因克隆。该蛋白片段是从稻瘟菌的培养滤液中分离出来的,并通过从头测序进行了鉴定。根据序列比较和 PCR 扩增,克隆了编码该激发子的基因 mohrip2。该 459 个碱基对的基因编码一个 152 个氨基酸残基的多肽,包含一个 18 个氨基酸残基的信号肽,等电点为 4.72,表观分子量为 16 kDa。该假设蛋白 MoHrip2 在大肠杆菌中表达,重组蛋白和内源性蛋白均能引起烟草叶片坏死。除了酚类化合物的沉积和细胞外介质的碱化作用外,MoHrip2 还能诱导烟草细胞产生过氧化氢和一氧化氮的积累。此外,与对照幼苗相比,用 MoHrip2 处理的水稻幼苗对稻瘟病菌表现出明显的抗性。