In Situ Study of Metabolic Response of Arabidopsis thaliana Leaves to Salt Stress by Neutral Desorption-Extractive Electrospray Ionization Mass Spectrometry

Salt stress is one of the most common factors limiting plant cultivation. In this study, metabolic responses to salt stress in Arabidopsis thaliana (A. thaliana) leaves were analyzed in situ by neutral desorption-extractive electrospray ionization mass spectrometry (ND-EESI-MS) without any sample pretreatment. Metabolic changes of A. thaliana leaves were observed in response to salt stress conditions, including the levels of serine, glutamic acid, arginine, cinnamic acid, ferulic acid, caffeic acid, protocatechuic acid, epicatechin, morin, myricetin, apigravin, and β-cotonefuran. The content of serine increased under 50, 100, and 200 mM NaCl salt stress, reaching the highest level at 200 mM NaCl, but decreased under the maximum concentration of 300 mM NaCl. A similar phenomenon was observed for arginine, glutamic acid, cinnamic acid, caffeic acid, ferulic acid, and epicatechin, respectively, involved in the metabolic pathway of shikimate-phenylpropanoid. Both principal component analysis (PCA) and partial least-squares discrimination analysis (PLS-DA) showed that the salt stress treatment groups of the higher concentrations (200 and 300 mM) could be well distinguished from those of the lower concentrations (50 and 100 mM) and the control. Marker metabolites, like m/z 261 (apigravin) and m/z 305 (β-cotonefuran), were assistantly selected from the fingerprints by variable importance for the projection (VIP). Our results indicated the potential of the ND-EESI-MS method for the rapid recognition of metabolic conditions in plant leaves under salt stress.