UHPLC-MS/MS based target profiling of stress-induced phytohormones

Area of Science:

• Plant Science
• Metabolomics
• Biochemistry

Background:

• Plant hormones regulate growth and stress responses.
• Jasmonates (JAs) are key regulators involved in plant defense and development.
• Understanding JA interactions with other hormones like salicylic acid (SA), abscisic acid (ABA), and indole-3-acetic acid (IAA) is crucial.

Purpose of the Study:

• To develop an efficient single-step clean-up procedure for phytohormone enrichment from plant matrices.
• To enable sensitive quantitative analysis of phytohormones using hyphenated mass spectrometry.
• To simultaneously profile multiple stress-induced phytohormones in small plant samples.

Main Methods:

• Developed a rapid extraction protocol using cold 10% methanol for minute plant material (<20mg FW).
• Employed a one-step reversed-phase polymer-based solid-phase extraction for analyte enrichment.
• Paired the protocol with a validated ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method.

Main Results:

• Significantly reduced matrix effects and increased recovery of labile JA analytes.
• Successfully profiled sixteen stress-induced phytohormones, including JA, its precursors/derivatives, SA, ABA, and IAA.
• Enabled sensitive analysis of phytohormones from minute plant material samples.

Conclusions:

• The developed protocol offers a highly efficient and sensitive method for phytohormone profiling.
• This technique facilitates the study of complex phytohormone interactions under stress conditions.
• The method is valuable for analyzing small plant samples, aiding research in plant stress physiology and development.