Plant Hormonomics: Multiple Phytohormone Profiling by Targeted Metabolomics

Abstract

Phytohormones are physiologically important small molecules that play essential roles in intricate signaling networks that regulate diverse processes in plants. We present a method for the simultaneous targeted profiling of 101 phytohormone-related analytes from minute amounts of fresh plant material (less than 20 mg). Rapid and nonselective extraction, fast one-step sample purification, and extremely sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry enable concurrent quantification of the main phytohormone classes: cytokinins, auxins, brassinosteroids, gibberellins, jasmonates, salicylates, and abscisates. We validated this hormonomic approach in salt-stressed and control Arabidopsis () seedlings, quantifying a total of 43 endogenous compounds in both root and shoot samples. Subsequent multivariate statistical data processing and cross-validation with transcriptomic data highlighted the main hormone metabolites involved in plant adaptation to salt stress.

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“…Endogenous levels of plant hormones (JAs, GAs, and SA) were measured using 20 mg (fresh weight) of uncolonized poplar LRs and ECM, as well as L. bicolor FLM, as described by Šimura et al (). Five biological replicates per condition were produced.…”

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“…Šimura et al [68] also observed in salt-stressed root samples increased levels of abscisic acid, its oxidation products phaseic acid, dihydrophaseic acid, and jasmoic acid, too. In addition, different responses of gibberillinic acid derivatives, especially its active form GA4, and its transcriptomic data were found by Šimura et al [68].…”

“…In addition, different responses of gibberillinic acid derivatives, especially its active form GA4, and its transcriptomic data were found by Šimura et al [68]. In sum, this newly developed multiple parallel analysis of phytohormones, called "plant hormonomics" enables the real-time profiling of hormone networks of large collections of phytohormones, their precursors, transport forms and degradation products in single stressed or non-stressed samples [68,70].…”

“…Although, Šimura et al [68] were already able to quantify the main phytohormones in less than 20 mg of salt stressed Arabidopsis samples, special derivatisation reactions of the analytes prior to their analysis lead to a conceivable increased sensitivity and selectivity during MS analysis in the future [67,[70][71][72]. For example, the response/sensitivity during LC-ESI-MS/MS analysis of metabolites only present in very low concentrations in plants, like indole-3-acetic acid, a member of the auxin family, can be tremendously increased up to 200 times after methylation [71].…”

“…[67]; Novák et al [63]) during the last two decades, exclusively one method was described to characterize those physiologically important molecules in their network interactions in one single run [68]. Šimura et al [68] also observed in salt-stressed root samples increased levels of abscisic acid, its oxidation products phaseic acid, dihydrophaseic acid, and jasmoic acid, too.…”