Rapid and Sensitive Hormonal Profiling of Complex Plant Samples by Liquid Chromatography Coupled to Electrospray Ionization Tandem Mass Spectrometry

Overview

Journal Plant Methods

Publisher Biomed Central

Specialty Biology

Date 2011 Nov 22

PMID 22098763

Citations 119

Authors

Maren Muller

Sergi Munne-Bosch

Affiliations

Soon will be listed here.

Abstract

Background: Plant 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 (10-9 M to 10-6 M) and their different chemistries, the development of a high-throughput and comprehensive method for the determination of hormones is challenging.

Results: The 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.

Conclusions: This 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.

Citing Articles

Natural variation in MdNAC5 contributes to fruit firmness and ripening divergence in apple.

Liu L, Wang Y, Guo J, Han Z, Yu K, Song Y Hortic Res. 2025; 12(1):uhae284.

PMID: 39866962 PMC: 11758708. DOI: 10.1093/hr/uhae284.

Phytohormonal homeostasis, chloroplast stability, and heat shock transcription pathways related to the adaptability of creeping bentgrass species to heat stress.

Yang H, Yuan Y, Liu X, Du Y, Li Z Protoplasma. 2025; .

PMID: 39794516 DOI: 10.1007/s00709-024-02022-0.

Adaptation of potato cultivars to phosphorus variability and enhancement of phosphorus efficiency by Bacillus subtilis.

Chea L, Alhussein M, Karlovsky P, Pawelzik E, Naumann M BMC Plant Biol. 2024; 24(1):1176.

PMID: 39668377 PMC: 11636052. DOI: 10.1186/s12870-024-05868-x.

Interactive effects of Orobanche latisquama parasitism and drought stress in Salvia rosmarinus plants growing under Mediterranean field conditions.

Jene L, Masso-Rodriguez M, Munne-Bosch S Physiol Plant. 2024; 176(6):e14652.

PMID: 39641143 PMC: 11621997. DOI: 10.1111/ppl.14652.

Variability in strawberry tunnels impacts fruit quality and limits melatonin effects.

Arabia A, Pallares N, Munne-Bosch S, Munoz P J Sci Food Agric. 2024; 105(3):1745-1759.

PMID: 39394858 PMC: 11726618. DOI: 10.1002/jsfa.13951.

References

Chen W, Gai Y, Liu S, Wang R, Jiang X . Quantitative analysis of cytokinins in plants by high performance liquid chromatography: electronspray ionization ion trap mass spectrometry. J Integr Plant Biol. 2010; 52(10):925-32. DOI: 10.1111/j.1744-7909.2010.00989.x. View

Kojima M, Kamada-Nobusada T, Komatsu H, Takei K, Kuroha T, Mizutani M . Highly sensitive and high-throughput analysis of plant hormones using MS-probe modification and liquid chromatography-tandem mass spectrometry: an application for hormone profiling in Oryza sativa. Plant Cell Physiol. 2009; 50(7):1201-14. PMC: 2709547. DOI: 10.1093/pcp/pcp057. View

Goda H, Sawa S, Asami T, Fujioka S, Shimada Y, Yoshida S . Comprehensive comparison of auxin-regulated and brassinosteroid-regulated genes in Arabidopsis. Plant Physiol. 2004; 134(4):1555-73. PMC: 419831. DOI: 10.1104/pp.103.034736. View

Richards D, King K, Ait-Ali T, Harberd N . HOW GIBBERELLIN REGULATES PLANT GROWTH AND DEVELOPMENT: A Molecular Genetic Analysis of Gibberellin Signaling. Annu Rev Plant Physiol Plant Mol Biol. 2001; 52:67-88. DOI: 10.1146/annurev.arplant.52.1.67. View

Vlot A, Dempsey D, Klessig D . Salicylic Acid, a multifaceted hormone to combat disease. Annu Rev Phytopathol. 2009; 47:177-206. DOI: 10.1146/annurev.phyto.050908.135202. View

Wi S, Park K . Antisense expression of carnation cDNA encoding ACC synthase or ACC oxidase enhances polyamine content and abiotic stress tolerance in transgenic tobacco plants. Mol Cells. 2002; 13(2):209-20. View

Muller A, Duchting P, Weiler E . A multiplex GC-MS/MS technique for the sensitive and quantitative single-run analysis of acidic phytohormones and related compounds, and its application to Arabidopsis thaliana. Planta. 2002; 216(1):44-56. DOI: 10.1007/s00425-002-0866-6. View

Wang D, Pajerowska-Mukhtar K, Culler A, Dong X . Salicylic acid inhibits pathogen growth in plants through repression of the auxin signaling pathway. Curr Biol. 2007; 17(20):1784-90. DOI: 10.1016/j.cub.2007.09.025. View

Munns R . Comparative physiology of salt and water stress. Plant Cell Environ. 2002; 25(2):239-250. DOI: 10.1046/j.0016-8025.2001.00808.x. View

10.

Raghavendra A, Gonugunta V, Christmann A, Grill E . ABA perception and signalling. Trends Plant Sci. 2010; 15(7):395-401. DOI: 10.1016/j.tplants.2010.04.006. View

11.

Pic E, Teyssendier de la Serve B, Tardieu F, Turc O . Leaf senescence induced by mild water deficit follows the same sequence of macroscopic, biochemical, and molecular events as monocarpic senescence in pea. Plant Physiol. 2002; 128(1):236-46. PMC: 148985. View

12.

Wilkinson S, Davies W . Drought, ozone, ABA and ethylene: new insights from cell to plant to community. Plant Cell Environ. 2009; 33(4):510-25. DOI: 10.1111/j.1365-3040.2009.02052.x. View

13.

Ghanem M, Albacete A, Martinez-Andujar C, Acosta M, Romero-Aranda R, Dodd I . Hormonal changes during salinity-induced leaf senescence in tomato (Solanum lycopersicum L.). J Exp Bot. 2008; 59(11):3039-50. PMC: 2504355. DOI: 10.1093/jxb/ern153. View

14.

Chiwocha S, Abrams S, Ambrose S, Cutler A, Loewen M, Ross A . A method for profiling classes of plant hormones and their metabolites using liquid chromatography-electrospray ionization tandem mass spectrometry: an analysis of hormone regulation of thermodormancy of lettuce (Lactuca sativa L.) seeds. Plant J. 2003; 35(3):405-17. DOI: 10.1046/j.1365-313x.2003.01800.x. View

15.

Izumi Y, Okazawa A, Bamba T, Kobayashi A, Fukusaki E . Development of a method for comprehensive and quantitative analysis of plant hormones by highly sensitive nanoflow liquid chromatography-electrospray ionization-ion trap mass spectrometry. Anal Chim Acta. 2009; 648(2):215-25. DOI: 10.1016/j.aca.2009.07.001. View

16.

Segarra G, Jauregui O, Casanova E, Trillas I . Simultaneous quantitative LC-ESI-MS/MS analyses of salicylic acid and jasmonic acid in crude extracts of Cucumis sativus under biotic stress. Phytochemistry. 2006; 67(4):395-401. DOI: 10.1016/j.phytochem.2005.11.017. View

17.

Nemhauser J, Mockler T, Chory J . Interdependency of brassinosteroid and auxin signaling in Arabidopsis. PLoS Biol. 2004; 2(9):E258. PMC: 509407. DOI: 10.1371/journal.pbio.0020258. View

18.

Pan X, Wang X . Profiling of plant hormones by mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci. 2009; 877(26):2806-13. DOI: 10.1016/j.jchromb.2009.04.024. View

19.

Dun E, Brewer P, Beveridge C . Strigolactones: discovery of the elusive shoot branching hormone. Trends Plant Sci. 2009; 14(7):364-72. DOI: 10.1016/j.tplants.2009.04.003. View

20.

Pan X, Welti R, Wang X . Simultaneous quantification of major phytohormones and related compounds in crude plant extracts by liquid chromatography-electrospray tandem mass spectrometry. Phytochemistry. 2008; 69(8):1773-81. DOI: 10.1016/j.phytochem.2008.02.008. View