Centre for Plant Metabolomics, Department of Biochemistry, University of Johannesburg, Auckland Park, Johannesburg, South Africa.
Department of Clinical Cancer Prevention, University of Texas MD Anderson Cancer Centre, 1515 Holcombe Blvd., Houston, TX, 77030, USA.
Sci Rep. 2019 Mar 1;9(1):3290. doi: 10.1038/s41598-019-38692-7.
Metabolomics has emerged as a powerful approach to comprehensively interrogate cellular biochemistry. As such, we applied an untargeted liquid chromatography-mass spectrometry metabolomic strategy to elucidate metabolome changes in the anthracnose-causing hemibiotrophic sorghum pathogen, Colletotrichum sublineolum. An in vitro batch culture study model with different carbon sources, glucose, arabinose and rhamnose, were used to support fungal growth over a period of twelve days. Metabolites representing the intracellular and extracellular (secreted) metabolomes were extracted with methanol and subjected to LC-MS analyses. Chemometric modelling revealed a metabolic variation trajectory, comprising three distinct stages that metabolically describe the adaptation of the fungus to diminishing nutrients. Selected marker gene expression indicated stage one (0-3 d.p.i) as corresponding to the early logarithmic phase. Stage two can be interpreted as an intermediate transitionary stage with stage three corresponding to the stationary phase (9-12 d.p.i). Stage one was characterised by up-regulation of endo-metabolites such as ferricrocin, fatty acids and flavone-conjugates, while stage three was characterised by the secretion of phytotoxins, including colletotrichin and colletotric acid. Ultimately, results from our in vitro model reveal previously unknown insights into the dynamic aspects of metabolome reprogramming in the growth phases of Colletotrichum spp as determined by nutrients obtainable from plant cell walls.
代谢组学已成为全面研究细胞生物化学的有力方法。因此，我们应用非靶向液相色谱-质谱代谢组学策略来阐明炭疽病引起的半活体高粱病原体，炭疽菌，的代谢组变化。使用不同的碳源，葡萄糖，阿拉伯糖和鼠李糖，进行体外分批培养研究模型，以支持真菌在十二天的时间内生长。用甲醇提取代表细胞内和细胞外（分泌）代谢组的代谢物，并进行 LC-MS 分析。化学计量模型揭示了代谢变化轨迹，包括三个不同的阶段，这些阶段在代谢上描述了真菌对营养物质减少的适应。选定的标记基因表达表明第一阶段（0-3 d.p.i）对应于早期对数期。第二阶段可以解释为中间过渡阶段，第三阶段对应于静止期（9-12 d.p.i）。第一阶段的特征是内代谢物如 ferricrocin、脂肪酸和黄酮缀合物的上调，而第三阶段的特征是植物细胞壁可利用的营养物质分泌出包括炭疽菌素和炭疽酸在内的植物毒素。最终，我们的体外模型的结果揭示了以前未知的关于 Colletotrichum spp 生长阶段代谢组重编程的动态方面的见解。
