Standardizing Methods for Shoot Metabolomics in Mycorrhizal Plants.

With the latest advances in analytical techniques based on liquid chromatography (LC) coupled with mass spectrometry (MS), knowledge of plant metabolomics has risen exponentially in recent years. The study of metabolomic changes associated with mycorrhizal symbiosis interacting with different environmental situations exemplifies the expansion of knowledge in this field. In the present chapter, we aim to provide a standard procedure for the analysis of shoot metabolites using liquid chromatography coupled with high-resolution mass spectrometry. The provided information includes an extraction buffer of compromised polarity, as well as LC and MS conditions suitable for a general characterization of secondary metabolites from mycorrhizal plants. These conditions may require further adaptation in case a lipidomic or highly polar compound analysis is required or when a different instrumentation is used. In addition, we provide a protocol for a preliminary bioinformatic analysis of the identified features using public non-proprietary software, which, combined with the construction of pure standard libraries, can yield a powerful tool for the identification and semi-quantitative analysis of hundreds of secondary metabolites from mycorrhizal plants.