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Proteomic Analysis of Phytophthora infestans Reveals the Importance of Cell Wall Proteins in Pathogenicity

机译:疫霉疫霉的蛋白质组学分析揭示了细胞壁蛋白在致病性中的重要性

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摘要

The oomycete Phytophthora infestans is the most harmful pathogen of potato. It causes the disease late blight, which generates increased yearly costs of up to one billion euro in the EU alone and is difficult to control. We have performed a large-scale quantitative proteomics study of six P. infestans life stages with the aim to identify proteins that change in abundance during development, with a focus on preinfectious life stages. Over 10 000 peptides from 2061 proteins were analyzed. We identified several abundance profiles of proteins that were up- or downregulated in different combinations of life stages. One of these profiles contained 59 proteins that were more abundant in germinated cysts and appressoria. A large majority of these proteins were not previously recognized as being appressorial proteins or involved in the infection process. Among those are proteins with putative roles in transport, amino acid metabolism, pathogenicity (including one RXLR effector) and cell wall structure modification. We analyzed the expression of the genes encoding nine of these proteins using RT-qPCR and found an increase in transcript levels during disease progression, in agreement with the hypothesis that these proteins are important in early infection. Among the nine proteins was a group involved in cell wall structure modification and adhesion, including three closely related, uncharacterized proteins encoded by PITG_01131, PITG_01132, and PITG_16135, here denoted Piacwp1–3. Transient silencing of these genes resulted in reduced severity of infection, indicating that these proteins are important for pathogenicity. Our results contribute to further insight into P. infestans biology, and indicate processes that might be relevant for the pathogen while preparing for host cell penetration and during infection. The mass spectrometry data have been deposited to ProteomeXchange via the PRIDE partner repository with the data set identifier PXD002446.
机译:卵菌疫霉菌是马铃薯中最有害的病原体。它会导致疾病晚疫病,仅在欧盟每年就增加高达10亿欧元的成本,而且很难控制。我们已经对六个致病疫霉的生命阶段进行了大规模的定量蛋白质组学研究,目的是鉴定在发育过程中蛋白质丰度变化的蛋白质,重点是感染前的生命阶段。分析了来自2061种蛋白质的10000多条肽。我们确定了在生命阶段的不同组合中被上调或下调的几种蛋白质丰度曲线。这些图谱之一包含59种蛋白质,这些蛋白质在发芽的囊肿和黏膜中更为丰富。这些蛋白质中的绝大多数以前未被认为是附属蛋白质或不参与感染过程。其中包括在运输,氨基酸代谢,致病性(包括一种RXLR效应子)和细胞壁结构修饰中具有推定作用的蛋白质。我们使用RT-qPCR分析了编码其中9种蛋白质的基因的表达,发现在疾病进展过程中转录水平有所增加,这与这些蛋白质在早期感染中很重要的假设相符。在这9种蛋白质中,有一组参与细胞壁结构的修饰和粘附,包括由PITG_01131,PITG_01132和PITG_16135编码的三种紧密相关,未表征的蛋白质,在这里称为Piacwp1-3。这些基因的瞬时沉默导致感染严重程度降低,表明这些蛋白质对于致病性很重要。我们的结果有助于进一步了解致病疫霉生物学,并指出在准备宿主细胞穿透和感染期间可能与病原体相关的过程。质谱数据已通过PRIDE合作伙伴存储库以数据集标识符PXD002446存放到ProteomeXchange。

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