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Rapid LC-MS-based metabolomics method to study the Fusarium infection of barley

机译:基于LC-MS的快速代谢组学方法研究大麦镰刀菌感染

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Ultra high performance liquid chromatography with quadrupole/time-of-flight mass spectrometry was applied to evaluate the potential of nontarget metabolomic fingerprinting in order to distinguish Fusarium-infected and control barley samples. First, the sample extraction and instrumental conditions were optimized to obtain the broadest possible representation of polar/medium-polar compounds occurring in extracts obtained from barley grain samples. Next, metabolomic fingerprints of extracts obtained from nine barley varieties were acquired under ESI conditions in both positive and negativemode. Each variety of barley was tested in two variants: artificially infected by Fusarium culmorum at the beginning of heading and a control group (no infection). In addition, the dynamics of barley infection development was monitored using this approach. The experimental data were statistically evaluated by principal component analysis, hierarchical clustering analysis, and orthogonal partial least-squares discriminant analysis. The differentiation of barley in response to F. culmorum infection was feasible using this metabolomics-based method. Analysis in positive mode provided a higher number of molecular features as compared to that performed under negative mode setting. However, the analysis in negative mode permitted the detection of deoxynivalenol and deoxynivalenol-3-glucoside considered as resistance-indicator metabolites in barley.
机译:采用四极杆/飞行时间质谱的超高效液相色谱法来评估非目标代谢组学指纹分析的潜力,以区分感染镰刀菌的大麦样品和对照大麦样品。首先,对样品提取和仪器条件进行了优化,以获得从大麦籽粒样品中提取的极性/中等极性化合物的最广泛代表。接下来,在ESI条件下,以正模式和负模式获取了从9个大麦品种获得的提取物的代谢组学指纹。每个大麦品种均以两种变体进行了测试:抽穗初期由镰刀菌人工感染和对照组(无感染)。此外,使用这种方法监测了大麦感染发生的动态。通过主成分分析,层次聚类分析和正交偏最小二乘判别分析对实验数据进行统计学评估。使用这种基于代谢组学的方法,可以对大麦对镰刀菌的感染做出区分。与在负模式下进行的分析相比,以正模式进行的分析提供了更多的分子特征。但是,以阴性模式进行分析可以检测出被认为是大麦中抗药性指标代谢产物的脱氧雪腐烯醇和脱氧雪腐烯醇-3-葡萄糖苷。

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