title: UHPLC-MS/MS based target profiling of stress-induced phytohormones.
abstract: Stress-induced changes in phytohormone metabolite profiles have rapid effects on plant metabolic activity and growth. The jasmonates (JAs) are a group of fatty acid-derived stress response regulators with roles in numerous developmental processes. To elucidate their dual regulatory effects, which overlap with those of other important defence-signalling plant hormones such as salicylic acid (SA), abscisic acid (ABA) and indole-3-acetic acid (IAA), we have developed a highly efficient single-step clean-up procedure for their enrichment from complex plant matrices that enables their sensitive quantitative analysis using hyphenated mass spectrometry technique. The rapid extraction of minute quantities of plant material (less than 20mg fresh weight, FW) into cold 10% methanol followed by one-step reversed-phase polymer-based solid phase extraction significantly reduced matrix effects and increased the recovery of labile JA analytes. This extraction and purification protocol was paired with a highly sensitive and validated ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method and used to simultaneously profile sixteen stress-induced phytohormones in minute plant material samples, including endogenous JA, several of its biosynthetic precursors and derivatives, as well as SA, ABA and IAA.
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany AS CR & Palacký University, Šlechtitelů 11, CZ-78371 Olomouc, Czech Republic.
authors.forename: Kristýna
authors.initials: K
authors.lastname: Floková
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany AS CR & Palacký University, Šlechtitelů 11, CZ-78371 Olomouc, Czech Republic.
authors.forename: Danuše
authors.initials: D
authors.lastname: Tarkowská
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany AS CR & Palacký University, Šlechtitelů 11, CZ-78371 Olomouc, Czech Republic.
authors.forename: Otto
authors.initials: O
authors.lastname: Miersch
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany AS CR & Palacký University, Šlechtitelů 11, CZ-78371 Olomouc, Czech Republic.
authors.forename: Miroslav
authors.initials: M
authors.lastname: Strnad
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany AS CR & Palacký University, Šlechtitelů 11, CZ-78371 Olomouc, Czech Republic.
authors.forename: Claus
authors.initials: C
authors.lastname: Wasternack
authors.affiliation: Laboratory of Growth Regulators, Centre of the Region Haná for Biotechnological and Agricultural Research, Institute of Experimental Botany AS CR & Palacký University, Šlechtitelů 11, CZ-78371 Olomouc, Czech Republic. Electronic address: novako@ueb.cas.cz.
authors.forename: Ondřej
authors.initials: O
authors.lastname: Novák
mesh_terms: D000040:Abscisic Acid / Q000032:analysis / Q000737:chemistry; D002853:Chromatography, Liquid; D003517:Cyclopentanes / Q000032:analysis / Q000737:chemistry; D005227:Fatty Acids / Q000032:analysis / Q000737:chemistry; D007210:Indoleacetic Acids / Q000032:analysis; D015394:Molecular Structure; D054883:Oxylipins / Q000032:analysis / Q000737:chemistry; D010937:Plant Growth Regulators / Q000737:chemistry / Q000302:isolation & purification* / Q000502:physiology*; D018515:Plant Leaves / Q000737:chemistry; D020156:Salicylic Acid / Q000032:analysis / Q000737:chemistry; D052616:Solid Phase Extraction; D013312:Stress, Physiological; D053719:Tandem Mass Spectrometry
chemical_list: D003517:Cyclopentanes; D005227:Fatty Acids; D007210:Indoleacetic Acids; D054883:Oxylipins; D010937:Plant Growth Regulators; C011006:jasmonic acid; C030737:indoleacetic acid; D000040:Abscisic Acid; D020156:Salicylic Acid
keywords: Abscisic acid; Arabidopsis thaliana; Indole-3-acetic acid; Jasmonates; Salicylic acid; Solid-phase extraction (SPE); Stress-induced phytohormones; Tandem mass spectrometry (MS/MS); Ultra-high performance liquid chromatography (UHPLC)