High-throughput interspecies profiling of acidic plant hormones using miniaturised sample processing

0564938 - ÚEB 2023 RIV GB eng J - Journal Article
Široká, Jitka - Brunoni, Federica - Pěnčík, Aleš - Mik, V. - Žukauskaitė, A. - Strnad, Miroslav - Novák, Ondřej - Floková, Kristýna
High-throughput interspecies profiling of acidic plant hormones using miniaturised sample processing.
Plant Methods. Roč. 18, č. 1 (2022), č. článku 122. E-ISSN 1746-4811
Institutional support: RVO:61389030
Keywords : 3D printing * Evolutionarily distant plant species * High-throughput * In-tip microSPE * Liquid chromatography * Mass spectrometry * Miniaturisation * Plant hormones
OECD category: Plant sciences, botany
Impact factor: 5.1, year: 2022
Method of publishing: Open access
https://doi.org/10.1186/s13007-022-00954-3

Background: Acidic phytohormones are small molecules controlling many physiological functions in plants. A comprehensive picture of their profiles including the active forms, precursors and metabolites provides an important insight into ongoing physiological processes and is essential for many biological studies performed on plants. Results: A high-throughput sample preparation method for liquid chromatography–tandem mass spectrometry determination of 25 acidic phytohormones classed as auxins, jasmonates, abscisates and salicylic acid was optimised. The method uses a small amount of plant tissue (less than 10 mg fresh weight) and acidic extraction in 1 mol/L formic acid in 10% aqueous methanol followed by miniaturised purification on reverse phase sorbent accommodated in pipette tips organised in a 3D printed 96-place interface, capable of processing 192 samples in one run. The method was evaluated in terms of process efficiency, recovery and matrix effects as well as establishing validation parameters such as accuracy and precision. The applicability of the method in relation to the amounts of sample collected from distantly related plant species was evaluated and the results for phytohormone profiles are discussed in the context of literature reports. Conclusion: The method developed enables high-throughput profiling of acidic phytohormones with minute amounts of plant material, and it is suitable for large scale interspecies studies.
Permanent Link: https://hdl.handle.net/11104/0336518