Identification of Bremia lactucae and Oidium neolycopersici proteins extracted for intact spore MALDI mass spectrometric biotyping

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• 作者：Jana Beinhauer ; René ; Lenobel ; Dmitry Loginov ; Ivo Chamrá ; d ; Pavel Řehulka ; Michaela Sedlá ; řová ; Martina Marchetti-Deschmann ; Gü ; nter Allmaier and Marek Šebela
• 刊名：ELECTROPHORESIS
• 出版年：2016
• 出版时间：November 2016
• 年：2016
• 卷：37
• 期：22
• 页码：2940-2952
• 全文大小：1476K
• ISSN：1522-2683

文摘

Several proteomic approaches were applied to identify protein markers providing typical signals during intact cell/spore (IC/IS) MALDI-TOF MS of two plant pathogens, namely Bremia lactucae (a downy mildew) and Oidium neolycopersici (a powdery mildew). First, proteins were extracted from intact spores of the microorganisms under conditions simulating their treatment prior to the mass spectrometric analysis. After a separation by electrophoresis and tryptic digestion, 198 and 140 proteins were identified in the B. lactucae and O. neolycopersici extracts, respectively. A large portion of them were found to be involved in the process of protein biosynthesis. For the first time, some proteins were assigned to characteristic signals in MS profiles of the investigated pathogens based on an agreement in the molecular mass. There were 9 and 10 proteins recognized, respectively, which could contribute significantly to the spectral patterns. These proteins were assigned tentatively to the following peaks in the MS profiles: (i) m/z 7828; 8593; 10 456; 11 312; 12 450; 12 763; 14 756 and 16 854 for B. lactucae; (ii) m/z 7709; 8895; 9504; 9952; 11 317; 14 082 and 14 839 for O. neolycopersici. We demonstrated the presence of ribosomal proteins and histones, which could be employed as markers in biotyping analyses for pathogen identification.