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Analysis of cell wall proteins regulated in stem of susceptible and resistant tomato species after inoculation with Ralstonia solanacearum: a proteomic approach

机译:番茄青枯雷尔菌接种后敏感和抗性番茄品种茎中调节的细胞壁蛋白分析:蛋白质组学方法

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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.
机译:蛋白质组学方法被用来阐明当番茄种被青枯病菌(Ralstonia solanacearum)接种时,在干细胞壁水平发生的分子相互作用。用盐缓冲液从纯化的细胞壁中提取抗性和易感植物在细菌接种前后的细胞壁蛋白,然后用2-D IEF / SDS-PAGE和3-D IEF / SDS / SDS-PAGE分离碱性蛋白。用胶体考马斯染色的凝胶显示出两种物种之间蛋白质斑点的丰度不同(抗药性中有八种蛋白质含量较高,易感性中有六种蛋白质含量较高)。此外,抗性(增加了七个蛋白质,减少了八个其他蛋白质)以及易感植物(五个蛋白质被升高,另外八个被抑制)中,响应细菌接种,蛋白质受到不同的调节。 MALDI-TOF / TOF MS和LC-ESI-IonTrap MS / MS的组合可鉴定那些蛋白质。植物通过增加两种物种中发病机理相关,其他防御相关和糖酵解蛋白的表达来响应病原体接种。然而,易感和抗氧化,应激相关的细胞壁代谢蛋白以及抗性品系中的能量代谢蛋白被抑制。预测比较分析的大多数蛋白质和其他随机选择的斑点,除一些经典的胞质蛋白质外,均具有分泌信号。

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