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In situ separation and visualization of isomeric auxin derivatives in Arabidopsis by ion mobility mass spectrometry imaging

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Abstract

In situ separation and visualization of synthetic and naturally occurring isomers from heterogeneous plant tissues, especially when they share similar molecular structures, are a challenging task. In this study, we combined the ion mobility separation with desorption electrospray ionization mass spectrometry imaging (DESI-IM-MSI) to achieve a direct separation and visualization of two synthetic auxin derivatives, auxinole and its structural isomer 4pTb-MeIAA, as well as endogenous auxins from Arabidopsis samples. Distinct distribution of these synthetic isomers and endogenous auxins in Arabidopsis primary roots and hypocotyls was achieved in the same imaging analysis from both individually treated and cotreated samples. We also observed putative metabolites of synthetic auxin derivatives, i.e. auxinole amino acid conjugates and hydrolysed 4pTb-MeIAA product — 4pTb-IAA, based on their unique drifting ion intensity patterns. Furthermore, DESI-IM-MSI-revealed abundance of endogenous auxins and synthetic isomers was validated by liquid chromatography–mass spectrometry (LC-MS). Our results demonstrate that DESI-IM-MSI could be used as a robust technique for detecting endogenous and exogenous isomers and provide a spatiotemporal evaluation of hormonomics profiles in plants.

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The data that supports the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors are thankful to Tomáš Pospíšil for valuable scientific discussions and Alexander Muck from Analytical Professional Services Group (Waters Corporation) for technical solutions.

Funding

This work was supported by the Internal Grant Agency of Palacký University Olomouc (IGA_PrF_2023_016, IGA_PrF_2023_031).

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Authors and Affiliations

  1. Laboratory of Growth Regulators, Institute of Experimental Botany, The Czech Academy of Sciences & Faculty of Science, Palacký University, Šlechtitelů 27, CZ-78371, Olomouc, Czech Republic

    Chao Zhang, Pavel Hladík, Ondřej Novák & Karel Doležal

  2. Department of Chemical Biology, Faculty of Science, Palacký University, Šlechtitelů 27, CZ-78371, Olomouc, Czech Republic

    Kristýna Bieleszová, Asta Žukauskaitė & Karel Doležal

  3. Department of Experimental Biology, Faculty of Science, Palacký University, Šlechtitelů 27, CZ-78371, Olomouc, Czech Republic

    Jiří Grúz

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  1. Chao Zhang
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Contributions

C.Z.: conceptualization; methodology; investigation; software; visualization; and writing, original draft. K.B.: investigation and formal analysis. A.Ž.: investigation; formal analysis; visualization; and writing, review and editing. P.H.: investigation and formal analysis. J.G.: resources. O.N.: funding acquisition; supervision; and writing, review and editing. K.D.: funding acquisition; supervision; and writing, review and editing.

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Correspondence to Chao Zhang or Karel Doležal.

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Zhang, C., Bieleszová, K., Žukauskaitė, A. et al. In situ separation and visualization of isomeric auxin derivatives in Arabidopsis by ion mobility mass spectrometry imaging. Anal Bioanal Chem 416, 125–139 (2024). https://doi.org/10.1007/s00216-023-04996-x

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