An UPLC-MS/MS method for highly sensitive high-throughput analysis of phytohormones in plant tissues

Plant Methods

Andrej Frolov²  Bettina Hause³  Alain Tissier³  Hagen Stellmach³  Anja Henning³  Mandy Fichtner³  Nick Bergau³  Vinzenz Handrick¹  Gerd Ulrich Balcke³ 

[1] Present address: Max Planck Institute for Chemical Ecology, Department of Biochemistry, Hans-Knoell-Str. 8, Jena, 07745, Germany;Faculty of Chemistry and Mineralogy, Institute of Bioanalytical Chemistry, Centre for Biotechnology and Biomedicine, Leipzig University, Deutscher Platz 5, Leipzig, 04103, Germany;Department of Cell and Metabolic Biology, Leibniz Institute of Plant Biochemistry, Weinberg 3, Halle (Saale), 06120, Germany

关键词: Wounding;    Plant stress response;    Electrospray ionization;    Quantification;    Solid phase extraction (SPE);    LC-MS/MS;    Jasmonates;    Phytohormones;   

Others  :  821271
DOI  :  10.1186/1746-4811-8-47

 received in 2012-09-05, accepted in 2012-11-12,  发布年份 2012

【 摘 要 】

Background

Phytohormones are the key metabolites participating in the regulation of multiple functions of plant organism. Among them, jasmonates, as well as abscisic and salicylic acids are responsible for triggering and modulating plant reactions targeted against pathogens and herbivores, as well as resistance to abiotic stress (drought, UV-irradiation and mechanical wounding). These factors induce dramatic changes in phytohormone biosynthesis and transport leading to rapid local and systemic stress responses. Understanding of underlying mechanisms is of principle interest for scientists working in various areas of plant biology. However, highly sensitive, precise and high-throughput methods for quantification of these phytohormones in small samples of plant tissues are still missing.

Results

Here we present an LC-MS/MS method for fast and highly sensitive determination of jasmonates, abscisic and salicylic acids. A single-step sample preparation procedure based on mixed-mode solid phase extraction was efficiently combined with essential improvements in mobile phase composition yielding higher efficiency of chromatographic separation and MS-sensitivity. This strategy resulted in dramatic increase in overall sensitivity, allowing successful determination of phytohormones in small (less than 50 mg of fresh weight) tissue samples. The method was completely validated in terms of analyte recovery, sensitivity, linearity and precision. Additionally, it was cross-validated with a well-established GC-MS-based procedure and its applicability to a variety of plant species and organs was verified.

Conclusion

The method can be applied for the analyses of target phytohormones in small tissue samples obtained from any plant species and/or plant part relying on any commercially available (even less sensitive) tandem mass spectrometry instrumentation.

【 授权许可】

   
2012 Balcke et al.; licensee BioMed Central Ltd.

【 图 表 】

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