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Plant Methods

, 7:37

First Online: 18 November 2011Received: 12 July 2011Accepted: 18 November 2011


BackgroundPlant hormones play a pivotal role in several physiological processes during a plant-s life cycle, from germination to senescence, and the determination of endogenous concentrations of hormones is essential to elucidate the role of a particular hormone in any physiological process. Availability of a sensitive and rapid method to quantify multiple classes of hormones simultaneously will greatly facilitate the investigation of signaling networks in controlling specific developmental pathways and physiological responses. Due to the presence of hormones at very low concentrations in plant tissues 10 M to 10 M and their different chemistries, the development of a high-throughput and comprehensive method for the determination of hormones is challenging.

ResultsThe present work reports a rapid, specific and sensitive method using ultrahigh-performance liquid chromatography coupled to electrospray ionization tandem spectrometry UPLC-ESI-MS-MS to analyze quantitatively the major hormones found in plant tissues within six minutes, including auxins, cytokinins, gibberellins, abscisic acid, 1-amino-cyclopropane-1-carboxyic acid the ethylene precursor, jasmonic acid and salicylic acid. Sample preparation, extraction procedures and UPLC-MS-MS conditions were optimized for the determination of all plant hormones and are summarized in a schematic extraction diagram for the analysis of small amounts of plant material without time-consuming additional steps such as purification, sample drying or re-suspension.

ConclusionsThis new method is applicable to the analysis of dynamic changes in endogenous concentrations of hormones to study plant developmental processes or plant responses to biotic and abiotic stresses in complex tissues. An example is shown in which a hormone profiling is obtained from leaves of plants exposed to salt stress in the aromatic plant, Rosmarinus officinalis.

KeywordsUPLC-ESI-MS-MS Phytohormones Auxins Abscisic acid Cytokinins Gibberellins Salicylic acid Jasmonic acid 1-amino-cyclopropane-1-carboxyic acid Rosmarinus officinalis AbbreviationsABAabscisic acid

ACC1-amino-cyclopropane-1-carboxyic acid

CEcollision energy: CXP: collision cell exit potential


DHZRdihydrozeatin riboside

DPdeclustering potencial

EPentrance potential

ESI-MS-MSelectrospray ionization tandem mass spectrometry

FPfocusing potential


IAAíndole-3-acetic acid

2iPisopentenyladenine: IPA: isopentenyladenosine

JAjasmonic acid

LODlimit of detection

LOQlimit of quantification

MRMmultiple reaction monitoring

OPDA12-oxo-phytodienoic acid

RDSrelative deviation standard

SAsalicylic acid

UPLCultrahigh performance liquid chromatography


ZRzeatin riboside.

Electronic supplementary materialThe online version of this article doi:10.1186-1746-4811-7-37 contains supplementary material, which is available to authorized users.

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Author: Maren Müller - Sergi Munné-Bosch

Source: https://link.springer.com/

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