Villela-Dias Cristiano, Camillo Luciana R, de Oliveira Guilherme A P, Sena Jamilly A L, Santiago André S, de Sousa Sanderson T P, Mendes Juliano S, Pirovani Carlos P, Alvim Fátima C, Costa Marcio G C
Laboratório de Proteômica, Centro de Biotecnologia e Genética, Universidade Estadual de Santa Cruz, Ilhéus, Bahia, Brazil; Mars Center for Cocoa Science, CP 55, Itajuipe, Bahia, Brazil.
Physiol Plant. 2014 Jan;150(1):1-17. doi: 10.1111/ppl.12061. Epub 2013 May 16.
NEP1 (necrosis- and ethylene-inducing peptide 1)-like proteins (NLPs) have been identified in a variety of taxonomically unrelated plant pathogens and share a common characteristic of inducing responses of plant defense and cell death in dicotyledonous plants. Even though some aspects of NLP action have been well characterized, nothing is known about the global range of modifications in proteome and metabolome of NLP-treated plant cells. Here, using both proteomic and metabolomic approaches we were able to identify the global molecular and biochemical changes in cells of Nicotiana benthamiana elicited by short-term treatment with MpNEP2, a NLP of Moniliophthora perniciosa, the basidiomycete responsible for the witches' broom disease on cocoa (Theobroma cacao L.). Approximately 100 protein spots were collected from 2-DE gels in each proteome, with one-third showing more than twofold differences in the expression values. Fifty-three such proteins were identified by mass spectrometry (MS)/MS and mapped into specific metabolic pathways and cellular processes. Most MpNEP2 upregulated proteins are involved in nucleotide-binding function and oxidoreductase activity, whereas the downregulated proteins are mostly involved in glycolysis, response to stress and protein folding. Thirty metabolites were detected by gas spectrometry (GC)/MS and semi-quantified, of which eleven showed significant differences between the treatments, including proline, alanine, myo-inositol, ethylene, threonine and hydroxylamine. The global changes described affect the reduction-oxidation reactions, ATP biosynthesis and key signaling molecules as calcium and hydrogen peroxide. These findings will help creating a broader understanding of NLP-mediated cell death signaling in plants.
NEP1（坏死和乙烯诱导肽1）样蛋白（NLPs）已在多种分类学上无关的植物病原体中被鉴定出来，它们具有一个共同特征，即能诱导双子叶植物产生植物防御反应和细胞死亡。尽管NLP作用的某些方面已得到充分表征，但对于经NLP处理的植物细胞蛋白质组和代谢组的全局修饰范围却一无所知。在这里，我们使用蛋白质组学和代谢组学方法，得以鉴定出由巴西可可丛枝病菌（Moniliophthora perniciosa）的NLP——MpNEP2短期处理引发的本氏烟草细胞中的全局分子和生化变化。在每个蛋白质组中，从二维凝胶上收集了大约100个蛋白质斑点，其中三分之一的蛋白质斑点表达值显示出两倍以上的差异。通过串联质谱（MS）/ MS鉴定了53种此类蛋白质，并将其映射到特定的代谢途径和细胞过程中。大多数MpNEP2上调的蛋白质参与核苷酸结合功能和氧化还原酶活性，而下调的蛋白质大多参与糖酵解、应激反应和蛋白质折叠。通过气相色谱（GC）/ MS检测并半定量了30种代谢物，其中11种在处理之间显示出显著差异，包括脯氨酸、丙氨酸、肌醇、乙烯、苏氨酸和羟胺。所描述的全局变化影响氧化还原反应、ATP生物合成以及钙和过氧化氢等关键信号分子。这些发现将有助于更全面地理解植物中NLP介导的细胞死亡信号传导。
