NMR-based metabolomic analysis of plants

Abstract

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 CD(3)OD:KH(2)PO(4) buffer in D(2)O), NMR analysis (standard (1)H, J-resolved, (1)H-(1)H 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.

A metabolomic study on the biological effects of metal pollutions in oysters Crassostrea sikamea

“…A global analysis on the metabolome profile in a biological system can provide metabolic responses induced by environmental stressors (Fasulo et al, 2012;Kwon et al, 2012;Cappello et al, 2013aCappello et al, , 2013b. And it has been also widely applied in multiple areas such as food quality (Tarachiwin et al, 2008), ecotoxicology (Jones et al, 2008) and plant sciences (Kim et al, 2010). Cappello et al (2013aCappello et al ( , 2013b identified the metabolic responses Hg and PAHs in mussel Mytilus galloprovincialis caged in anthropogenic-impacted sites along the Augusta coastline (Sicily, Italy) and confirmed these mussels had suffered from adverse environmental condition.…”

NMR-based pharmacometabonomics: A new paradigm for personalised or precision medicine

“…Each of these steps has been well investigated for either NMR- [14][15][16][17][18][19][20] or mass spectrometry (MS)-based [21][22][23][24] approaches to metabolic profiling. The step that presents the most difficulties to investigators using either NMR-based or MS-based approaches is metabolite identification (see Section 2.4 below).…”

A metabolomic investigation of the effects of metal pollution in oysters Crassostrea hongkongensis

“…Therefore, metabolomics can detect differences in the metabolome profile in response to environmental stressors (Cappello et al, 2013;Fasulo et al, 2012;Kwon et al, 2012). It has also been widely applied in multiple areas such as food quality (Tarachiwin et al, 2008), ecotoxicology (Jones et al, 2008) and plant sciences (Kim et al, 2010). Cappello et al (2013) successfully identified metabolic responses to environmental pollution, primarily from exposure to Hg and polyaromatic hydrocarbons (PAHs), in the mussel Mytilus galloprovincialis caged in anthropogenic-impacted and reference sites along the Augusta coastline (Sicily, Italy).…”