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Metabolomics

, Volume 11, Issue 4, pp 991–997 | Cite as

Identification of the 12-oxojasmonoyl-isoleucine, a new intermediate of jasmonate metabolism in Arabidopsis, by combining chemical derivatization and LC–MS/MS analysis

  • Emilie Widemann
  • Thierry Heitz
  • Laurence Miesch
  • Michel Miesch
  • Clément Heinrich
  • Franck Pinot
  • Raphaël LuganEmail author
Original Article

Abstract

The Arabidopsis cytochrome P450 CYP94C1 is involved in the metabolism of jasmonates, a family of phytohormones implicated in plant development and responses to bioagressors. It has been shown to down-regulate the level of the active hormone jasmonoyl-isoleucine (JA-Ile) by catalyzing its successive oxidation into hydroxyl (12OH-JA-Ile) and carboxyl (12COOH-JA-Ile) derivatives. The systematic investigation of this enzymatic reaction in vitro reported here revealed the formation of a third oxidized product: UPLC–MS/MS data matched a possible aldehyde, an intermediate form between 12OH-JA-Ile and 12COOH-JA-Ile. To confirm the presence of an aldehyde group, the extracted metabolite from incubations were derivatized with methoxyamine, an agent known to target carbonyl functions specifically. UPLC–MS/MS analysis after derivatization confirmed the presence of 12-oxojasmonoyl-isoleucine (12CHO-JA-Ile). Methanolic extracts from Arabidopsis leaves were then investigated after wounding, a treatment that triggers the accumulation of JA-Ile and its oxidized derivatives. The 12CHO-JA-Ile produced a significant signal with the same order of magnitude than the other oxidized forms. This study illustrates the underestimated potential of chemical derivatization for rapid metabolite identification in modern metabolomics and enriches our knowledge of the jasmonates family.

Keywords

Liquid chromatography–mass spectrometry (LC–MS) Chemical derivatization Jasmonates Cytochrome P450 

Notes

Acknowledgments

This work was supported by a doctoral fellowship from the Ministère de l’Enseignement Supérieur et de la Recherche. It was also partially funded by the Agence Nationale de la Recherche grant ANR-12-BSV8-005. The UPLC–MS/MS instrument was co-financed by the Centre National de la Recherche Scientifique, the Université de Strasbourg, the Région Alsace, the Institut National de la Recherche Agronomique, and the Tepral Company.

Conflict of interest

All authors declare that they have no conflict of interest.

Compliance with ethical guidelines

This article does not contain any studies with human or animal subjects.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Emilie Widemann
    • 1
  • Thierry Heitz
    • 1
  • Laurence Miesch
    • 2
  • Michel Miesch
    • 2
  • Clément Heinrich
    • 2
  • Franck Pinot
    • 1
  • Raphaël Lugan
    • 1
    Email author
  1. 1.Institut de Biologie Moléculaire des Plantes, Unité Propre de Recherche 2357 CNRS, Université de StrasbourgStrasbourg CedexFrance
  2. 2.Laboratoire de Chimie Organique Synthétique, Institut de Chimie, Unité Mixte de Recherche 7177, Université de Strasbourg, CNRSStrasbourg CedexFrance

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