National Center for Soybean Improvement, National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, 210095, China.
Mol Biol Rep. 2011 Jan;38(1):511-21. doi: 10.1007/s11033-010-0135-x. Epub 2010 Apr 7.
Soybean mosaic virus (SMV) is one of the most serious virus diseases of soybean. However, little is known about the molecular basis of the soybean defense mechanism against this pathogen. We identified differentially expressed proteins in soybean leaves infected with SMV by proteomic approaches. Twenty-eight protein spots that showed ≥2-fold difference in intensity were identified between mock-inoculated and SMV-infected samples. Among them, 16 spots were upregulated and 12 spots were downregulated in the SMV-infected samples. We recovered 25 of the 28 differentially expressed proteins from two-dimensional electrophoresis (2-DE) gels. These spots were identified as 16 different proteins by Matrix assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) and tandem TOF/TOF MS, and were potentially involved in protein degradation, defense signal transfer, reactive oxygen, cell wall reinforcement, and energy and metabolism regulation. Gene expression analysis of 13 genes by quantitative real time polymerase chain reaction (qRT-PCR) showed that metabolism genes and photosynthesis genes were downregulated at all time points. One energy gene was downregulated, whereas another energy gene was upregulated at five of the six time points. The other interesting genes that were altered by SMV infection showed changes in transcription over time. This is the first extensive application of proteomics to the SMV-soybean interaction. These results contribute to a better understanding of the molecular basis of soybean's responses to SMV.
大豆花叶病毒(SMV)是大豆最严重的病毒病之一。然而,人们对大豆抵抗这种病原体的分子基础知之甚少。我们通过蛋白质组学方法鉴定了感染 SMV 的大豆叶片中的差异表达蛋白。在模拟接种和 SMV 感染的样品之间,有 28 个蛋白斑点的强度差异≥2 倍。其中,SMV 感染样品中 16 个斑点上调,12 个斑点下调。我们从二维电泳(2-DE)凝胶中回收了 28 个差异表达蛋白中的 25 个。这些斑点通过基质辅助激光解吸/电离飞行时间(MALDI-TOF)质谱(MS)和串联 TOF/TOF MS 鉴定为 16 种不同的蛋白质,它们可能参与蛋白质降解、防御信号转导、活性氧、细胞壁强化以及能量和代谢调节。通过实时定量聚合酶链反应(qRT-PCR)对 13 个基因的基因表达分析表明,代谢基因和光合作用基因在所有时间点都下调。一个能量基因下调,而另一个能量基因在六个时间点中的五个时间点上调。SMV 感染改变的其他有趣基因显示转录随时间的变化。这是蛋白质组学首次广泛应用于 SMV-大豆相互作用。这些结果有助于更好地理解大豆对 SMV 反应的分子基础。