Department of Molecular Genetics, Cereal Research Centre, Agriculture and Agrifood Canada, Winnipeg, MB, Canada.
Proteomics. 2013 Jun;13(12-13):1913-21. doi: 10.1002/pmic.201200446. Epub 2013 Apr 17.
To understand early events in plant-pathogen interactions, it is necessary to explore the pathogen secretome to identify secreted proteins that help orchestrate pathology. The secretome can be obtained from pathogens grown in vitro, and then characterized using standard proteomic approaches based on protein extraction and subsequent identification of tryptic peptides by LC-MS. A subset of the secretome is composed of proteins whose presence is required to initiate infection and their removal from the secretome would result in pathogens with reduced or no virulence. We present here comparative secretome from Fusarium graminearum. This filamentous fungus causes Fusarium head blight on wheat, a serious cereal disease found in many cereal-growing regions. Affected grain is contaminated with mycotoxins and cannot be used for food or feed. We used label-free quantitative MS to compare the secretomes of wild-type with two nonpathogenic deletion mutants of F. graminearum, Δtri6, and Δtri10. These mutations in mycotoxin-regulating transcription factors revealed a subset of 29 proteins whose relative abundance was affected in their secretomes, as measured by spectral counting. Proteins that decreased in abundance are potential candidate virulence factors and these included cell wall-degrading enzymes, metabolic enzymes, pathogenesis-related proteins, and proteins of unknown function.
为了了解植物-病原体相互作用的早期事件，有必要探索病原体的分泌组，以鉴定有助于调控病理学的分泌蛋白。可以从体外培养的病原体中获得分泌组，然后使用基于蛋白质提取的标准蛋白质组学方法对其进行表征，并通过 LC-MS 对酶切肽进行后续鉴定。分泌组的一部分由存在是为了启动感染所必需的蛋白质组成，如果从分泌组中去除这些蛋白质，会导致病原体的毒力降低或丧失。我们在这里展示来自禾谷镰刀菌的比较分泌组。这种丝状真菌在小麦上引起镰刀菌穗枯病，这是一种在许多谷物种植地区发现的严重谷类疾病。受感染的谷物被霉菌毒素污染，不能用于食品或饲料。我们使用无标记定量 MS 比较了野生型禾谷镰刀菌与两个非致病性缺失突变体Δtri6 和Δtri10 的分泌组。这些在调控霉菌毒素的转录因子中的突变揭示了一个由 29 种蛋白质组成的子集，它们的相对丰度在其分泌组中受到影响，这是通过光谱计数来衡量的。丰度降低的蛋白质可能是潜在的候选毒力因子，其中包括细胞壁降解酶、代谢酶、与发病机制相关的蛋白质和未知功能的蛋白质。
