Institute for Plant Diseases and Plant Protection, Faculty of Natural Sciences, Leibniz Universität Hannover, Herrenhäuser Str.2, 30419 Hannover, Germany.
Plant Mol Biol. 2010 Aug;73(6):643-58. doi: 10.1007/s11103-010-9646-z. Epub 2010 May 23.
Proteomics approach was used to elucidate the molecular interactions taking place at the stem cell wall level when tomato species were inoculated with Ralstonia solanacearum, a causative agent of bacterial wilt. Cell wall proteins from both resistant and susceptible plants before and after the bacterial inoculation were extracted from purified cell wall with salt buffers and separated with 2-D IEF/SDS-PAGE and with 3-D IEF/SDS/SDS-PAGE for basic proteins. The gels stained with colloidal Coomassie revealed varied abundance of protein spots between two species (eight proteins in higher abundance in resistant and six other in susceptible). Moreover, proteins were regulated differentially in response to bacterial inoculation in resistant (seven proteins increased and eight other decreased) as well as in susceptible plants (five proteins elevated and eight other suppressed). Combination of MALDI-TOF/TOF MS and LC-ESI-IonTrap MS/MS lead to the identification of those proteins. Plants responded to pathogen inoculation by elevating the expression of pathogenesis related, other defense related and glycolytic proteins in both species. However, cell wall metabolic proteins in susceptible, and antioxidant, stress related as well as energy metabolism proteins in resistant lines were suppressed. Most of the proteins of the comparative analysis and other randomly picked spots were predicted to have secretion signals except some classical cytosolic proteins.
采用蛋白质组学方法阐明了番茄物种接种青枯菌(细菌性萎蔫病的病原体)时,在干细胞壁水平上发生的分子相互作用。用盐缓冲液从纯化的细胞壁中提取接种前后抗性和敏感植物的细胞壁蛋白,并用 2-DIEF/SDS-PAGE 和 3-DIEF/SDS/SDS-PAGE 分离碱性蛋白。用胶体考马斯亮蓝染色的凝胶显示两种物种之间(抗性中 8 种蛋白丰度较高,敏感中 6 种蛋白丰度较低)蛋白斑点的丰度存在差异。此外,抗性(7 种蛋白增加,8 种蛋白减少)和敏感植物(5 种蛋白升高,8 种蛋白降低)对细菌接种的反应也存在差异。MALDI-TOF/TOF MS 和 LC-ESI-IonTrap MS/MS 的组合导致了这些蛋白质的鉴定。植物通过在两种物种中上调与发病机制相关、其他防御相关和糖酵解相关的蛋白来响应病原体接种。然而,敏感植物的细胞壁代谢蛋白以及抗性系中的抗氧化剂、应激相关和能量代谢蛋白被抑制。除了一些经典的胞质蛋白外,比较分析的大多数蛋白质和其他随机挑选的斑点都预测具有分泌信号。