Rampitsch Christof, Bykova Natalia V, McCallum Brent, Beimcik Eva, Ens Werner
Agriculture and Agrifood Canada, Winnipeg, MB, Canada.
Proteomics. 2006 Mar;6(6):1897-907. doi: 10.1002/pmic.200500351.
Wheat leaf rust is caused by the fungus Puccinia triticina. The genetics of resistance follows the gene-for-gene hypothesis, and thus the presence or absence of a single host resistance gene renders a plant resistant or susceptible to a leaf rust race bearing the corresponding avirulence gene. To investigate some of the changes in the proteomes of both host and pathogen during disease development, a susceptible line of wheat infected with a virulent race of leaf rust were compared to mock-inoculated wheat using 2-DE (with IEF pH 4-8) and MS. Up-regulated protein spots were excised and analyzed by MALDI-QqTOF MS/MS, followed by cross-species protein identification. Where possible MS/MS spectra were matched to homologous proteins in the NCBI database or to fungal ESTs encoding putative proteins. Searching was done using the MASCOT search engine. Remaining unmatched spectra were then sequenced de novo and queried against the NCBInr database using the BLAST and MS BLAST tools. A total of 32 consistently up-regulated proteins were examined from the gels representing the 9-day post-infection proteome in susceptible plants. Of these 7 are host proteins, 22 are fungal proteins of known or hypothetical function and 3 are unknown proteins of putative fungal origin.
小麦叶锈病由真菌小麦柄锈菌(Puccinia triticina)引起。抗性遗传学遵循基因对基因假说,因此单个宿主抗性基因的存在与否使植物对携带相应无毒基因的叶锈菌生理小种表现出抗性或易感性。为了研究病害发展过程中宿主和病原体蛋白质组的一些变化,使用二维电泳(IEF pH 4 - 8)和质谱法,将感染叶锈菌强毒生理小种的小麦感病品系与模拟接种的小麦进行比较。切除上调的蛋白质点并通过基质辅助激光解吸电离串联四极杆飞行时间质谱(MALDI - QqTOF MS/MS)进行分析,随后进行跨物种蛋白质鉴定。在可能的情况下,将串联质谱图与NCBI数据库中的同源蛋白质或编码假定蛋白质的真菌EST进行匹配。使用MASCOT搜索引擎进行搜索。然后对剩余的未匹配谱图进行从头测序,并使用BLAST和MS BLAST工具针对NCBInr数据库进行查询。从代表感病植物感染后9天蛋白质组的凝胶中总共检测了32个持续上调的蛋白质。其中7个是宿主蛋白质,22个是已知或假定功能的真菌蛋白质,3个是假定为真菌来源的未知蛋白质。