Nuclear magnetic resonance (NMR)-based metabolomics has many applications in plant science. Metabolomics can be used in functional genomics and to differentiate plants from different origin, or after different treatments. In this protocol, the following steps of plant metabolomics using NMR spectroscopy are described: sample preparation (freeze drying followed by extraction by ultrasonication with 1:1 CD3OD:KH2PO4 buffer in D2O), NMR analysis (standard 1H, J-resolved, 1H–1H correlation spectroscopy (COSY) and heteronuclear multiple bond correlation (HMBC)) and chemometric methods. The main advantage of NMR metabolomic analysis is the possibility of identifying metabolites by comparing NMR data with references or by structure elucidation using two-dimensional NMR. This protocol is particularly suited for the analysis of secondary metabolites such as phenolic compounds (usually abundant in plants), and for primary metabolites (e.g., sugars and amino acids). This procedure is rapid; it takes not more than 30 min for sample preparation (multiple parallel) and a further 10 min for NMR spectrum acquisition.
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We thank Ms. E.G. Wilson for reviewing the manuscript and providing helpful comments. We also thank Dr. A. Meissner, Mr. C. Erkelens and Mr. A.W.M. Lefeber for their kind help in setting up NMR parameters. This research has received funding from the European Community′s Seventh Framework Programme [FP7/2007-2013] under Grant Agreement No 217895.
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Kim, H., Choi, Y. & Verpoorte, R. NMR-based metabolomic analysis of plants. Nat Protoc 5, 536–549 (2010). https://doi.org/10.1038/nprot.2009.237
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