Pollier Jacob, Goossens Alain
Department of Plant Systems Biology, VIB, Gent, Belgium.
Methods Mol Biol. 2013;1011:277-86. doi: 10.1007/978-1-62703-414-2_22.
Plants accumulate an overwhelming variety of secondary metabolites that play important roles in defense and interaction of the plant with its environment. To investigate the dynamics of plant secondary metabolism, large-scale untargeted metabolite profiling (metabolomics) is mandatory. Here, we describe a detailed protocol for untargeted metabolite profiling in which methanol extracts of jasmonate-treated plant tissues are analyzed by reversed-phase liquid chromatography coupled to negative-ion electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (MS). By means of dedicated integration and alignment software, the relative abundance of thousands of mass peaks, corresponding to hundreds of compounds, is calculated, and mass peaks of which the area is significantly changed by jasmonate treatment are identified. Subsequently, the metabolites corresponding to the significantly changed peaks are tentatively annotated using the accurate mass prediction of the Fourier transform-MS and the generated MS/MS data. Via this method, compounds of medium polarity, such as glucosinolates, alkaloids, phenylpropanoids, flavonoids, polyamines, and saponins, can be analyzed.
植物积累了种类繁多的次生代谢产物，这些产物在植物的防御以及与环境的相互作用中发挥着重要作用。为了研究植物次生代谢的动态变化，大规模非靶向代谢物谱分析（代谢组学）是必不可少的。在此，我们描述了一种详细的非靶向代谢物谱分析方案，其中通过反相液相色谱与负离子电喷雾电离傅里叶变换离子回旋共振质谱（MS）联用，对茉莉酸处理的植物组织的甲醇提取物进行分析。借助专用的积分和比对软件，计算出对应数百种化合物的数千个质谱峰的相对丰度，并识别出经茉莉酸处理后面积发生显著变化的质谱峰。随后，利用傅里叶变换-MS的精确质量预测和生成的MS/MS数据，对与显著变化峰对应的代谢物进行初步注释。通过这种方法，可以分析中等极性的化合物，如芥子油苷、生物碱、苯丙烷类、黄酮类、多胺和皂苷。
