Advertisement

Mass Spectrometric Approaches to Study the Metabolism of Jasmonates: Biotransformation of Exogenously Supplemented Methyl Jasmonate by Cell Suspension Cultures of Moringa oleifera

  • Claude Y. Hamany Djande
  • Ntakadzeni E. Madala
  • Ian A. DuberyEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2085)

Abstract

Jasmonic acid (JA) and derivatives play a crucial role in plant adaptation to environmental stress. The exogenous application of methyl jasmonate (MeJA) results in activation of stress-related genes and subsequent production of secondary metabolites. This implies the biotransformation of the hormone into a more active form. In this study, Moringa oleifera cell suspension cultures were treated with 100 μM MeJA. Methanolic cell extracts were analyzed with reverse phase ultrahigh performance liquid chromatography (UHPLC) coupled to a quadrupole time-of-flight high definition mass spectrometer (QTOF–HDMS, with MSE data acquisition). Using a targeted approach for jasmonate profiling, extracted ion chromatograms were generated. MS fragmentation patterns were subsequently derived and molecular formulae calculated from the MS data of each ion. Furthermore, triple quadrupole (QqQ) MS, operated in selected reaction monitoring (SRM) mode, was employed to target masses of interest. In addition to MeJA and JA, three jasmonoyl-amino acids were annotated: jasmonoyl-valine (JA-Val), jasmonoyl-isoleucine/leucine (JA-Ile/Leu), and jasmonoyl-phenylalanine (JA-Phe), based on characteristic precursor and product ions. Furthermore, JA conjugated to a hexose was observed, as well as hydroxylated and carboxylated derivatives of JA-amino acid conjugates. The data point to active metabolism of the externally added MeJA by the M. oleifera cells through biotransformation and bioconversion reactions that can be investigated in depth using advanced mass spectrometric analyses.

Key words

Bioconversion Jasmonic acid Jasmonoyl-amino acids Mass spectrometry Methyl jasmonate Moringa oleifera 

Notes

Acknowledgments

The University of Johannesburg and the South African National Research Foundation are acknowledged for fellowship support to CHD and grant support to IAD [grant number 95818]. Dr. PA Steenkamp is thanked for UHPLC–MS analyses, Ms. N Ndolvu for assistance with LC/MS/MS analyses, and Dr. F Tugizimana for assistance with data analysis.

References

  1. 1.
    Okada K, Abe H, Arimura G (2015) Jasmonates induce both defence responses and communication in mono- and dicotyledonous plants. Plant Cell Physiol 56:16–27CrossRefGoogle Scholar
  2. 2.
    Yan Y, Borrego E, Kolomiets MV (2013) Jasmonate biosynthesis, perception and function in plant development and stress responses. In: Lipid metabolism, vol 14. InTechOpen, London, pp 456–466.  https://doi.org/10.5772/52675CrossRefGoogle Scholar
  3. 3.
    Schaller A, Stintzi A (2008) Jasmonate biosynthesis and signaling for induced plant defence against herbivory. In: Induced plant resistance to herbivory. Springer, Dordrecht, pp 349–366CrossRefGoogle Scholar
  4. 4.
    Wasternack C, Strnad M (2016) Jasmonate signaling in plant stress responses and development – active and inactive compounds. Nat Biotechnol 33:604–613Google Scholar
  5. 5.
    Glauser G, Grata E, Dubugnon L, Rudaz S, Farmer EE, Wolfender J (2008) Spatial and temporal dynamics of jasmonate synthesis and accumulation in Arabidopsis in response to wounding. J Biol Chem 283:16400–16407CrossRefGoogle Scholar
  6. 6.
    Caarls L, Elberse J, Awwanah M, Ludwig NR, Vries M, Zeilmaker T, Van Wees SC, Schuurink RC, Van den Ackerveken G (2017) Arabidopsis jasmonate-induced oxygenases down-regulate plant immunity by hydroxylation and inactivation of the hormone jasmonic acid. Proc Natl Acad Sci 114:6399–6393CrossRefGoogle Scholar
  7. 7.
    Song S, Qi T, Wasternack C, Xie D (2014) Jasmonate signaling and crosstalk with gibberellin and ethylene. Curr Opin Plant Biol 21:112–119CrossRefGoogle Scholar
  8. 8.
    Widemann E, Grausem B, Renault H, Pineau E, Heinrich C, Lugan R, Ullmann P, Miesch L, Aubert Y, Miesch M, Heitz T (2015) Sequential oxidation of jasmonoyl-phenylalanine and jasmonoyl-isoleucine by multiple cytochrome P450 of the CYP94 family through newly identified aldehyde intermediates. Phytochemistry 117:388–399CrossRefGoogle Scholar
  9. 9.
    Woldemariam MG, Onkokesung N, Baldwin IT, Galis I (2012) Jasmonoyl-L-isoleucine hydrolase 1 (JIH1) regulates jasmonoyl-L-isoleucine levels and attenuates plant defences against herbivores. Plant J 1:758–767CrossRefGoogle Scholar
  10. 10.
    Koo AJ (2018) Metabolism of the plant hormone jasmonate: a sentinel for tissue damage and master regulator of stress response. Phytochem Rev 17.  https://doi.org/10.1007/s11101-017-9510-8CrossRefGoogle Scholar
  11. 11.
    Zhang T, Poudel AN, Jewell JB, Kitaoka N, Staswick P, Matsuura H, Koo AJ (2016) Hormone crosstalk in wound stress response: wound inducible amidohydrolases can simultaneously regulate jasmonate and auxin homeostasis in Arabidopsis thaliana. J Exp Bot 67:2107–2120CrossRefGoogle Scholar
  12. 12.
    Cimini S, Ronci MB, Barizza E, de Pinto MC, Locato V, Schiavo FL, De Gara L (2018) Plant cell cultures as model systems to study programmed cell death. In: Plant programmed cell death. Humana Press, New York, NY, pp 173–186CrossRefGoogle Scholar
  13. 13.
    James JT, Tugizimana F, Steenkamp PA, Dubery IA (2013) Metabolomic analysis of methyl jasmonate-induced triterpenoid production in the medicinal herb, Centella asiatica (L.) Urban. Molecules 18:4267–4281CrossRefGoogle Scholar
  14. 14.
    Mhlongo MI, Steenkamp PA, Piater LA, Madala NE, Dubery IA (2016) Profiling of altered metabolomic states in Nicotiana tabacum cells induced by priming agents. Front Plant Sci 7:1527CrossRefGoogle Scholar
  15. 15.
    Hamany Djande CY, Piater LA, Steenkamp PA, Madala NE, Dubery IA (2018) Differential extraction of phytochemicals from the multipurpose tree, Moringa oleifera, using green extraction solvents. S Afr J Bot 115:81–89CrossRefGoogle Scholar
  16. 16.
    Liu X, Yang Y, Lin W, Tong J, Huang Z, Xiao LT (2010) Determination of both jasmonic acid and methyl jasmonate in plant samples by liquid chromatography tandem mass spectrometry. Chinese Sci Bul 55:2231–2235CrossRefGoogle Scholar
  17. 17.
    Balcke GU, Handrick Bergau N, Fichtner N, Henning A et al (2012) An UPLC-MS/MS method for highly sensitive high-throughput analysis of phytohormones in plant tissues. Plant Methods 8:47CrossRefGoogle Scholar
  18. 18.
    Riet KB, Ndlovu N, Piater LA, Dubery IA (2016) Simultaneous analysis of defense-related phytohormones in Arabidopsis thaliana responding to fungal infection. Applic Plant Sci 4:1600013CrossRefGoogle Scholar
  19. 19.
    Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  20. 20.
    Wasternack C, Feussner I (2018) The oxylipin pathways: biochemistry and function. Annu Rev Plant Biol 69:363–386CrossRefGoogle Scholar
  21. 21.
    Göbel C, Feussner I (2009) Phytochemistry methods for the analysis of oxylipins in plants. Phytochemistry 70:1485–1503CrossRefGoogle Scholar
  22. 22.
    Eng F, Haroth S, Feussner K, Meldau D, Rekhter D, Ischebeck T, Brodhun F, Feussner I (2016) Optimized jasmonic acid production by Lasiodiplodia theobromae reveals formation of valuable plant secondary metabolites. PlosOne 11.  https://doi.org/10.1371/journal.pone.0167627CrossRefGoogle Scholar
  23. 23.
    Guranowski A, Miersch O, Staswick PE, Suza W, Wasternack C (2007) Substrate specificity and products of side-reactions catalyzed by jasmonate:amino acid synthetase (JAR1). FEBS Lett 581:815–820CrossRefGoogle Scholar
  24. 24.
    Staswick PE, Tiryaki I (2004) The oxylipin signal jasmonic acid is activated by an enzyme that conjugates it to isoleucine in Arabidopsis. Plant Cell 16:2117–2127CrossRefGoogle Scholar
  25. 25.
    Sumner LW, Amberg A, Barrett D, Beale MH, Beger R et al (2007) Proposed minimum reporting standards for chemical analysis: Chemical Analysis Working Group (CAWG) Metabolomics Standards Initiative (MSI). Metabolomics 3:211–221CrossRefGoogle Scholar
  26. 26.
    Han Y, Bai Y, Xiao Y, Du F, Liang Y, Tan Z, Zhao M, Liu H (2011) Simultaneous discrimination of jasmonic acid stereoisomers by CE-QTOF-MS employing the partial filling technique. Electrophoresis 32:2693–2699CrossRefGoogle Scholar
  27. 27.
    Matencio A, Bermejo-Gimeno MJ, Garcia-Carmona F, Lopez-Nicolas JM (2017) Separating and identifying the four stereoisomers of methy jasmonate by RP-HPLC and using cyclodextrins in a novel way. Phytochem Anal 28:151–158CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Claude Y. Hamany Djande
    • 1
  • Ntakadzeni E. Madala
    • 1
  • Ian A. Dubery
    • 1
    Email author
  1. 1.Department of Biochemistry, Centre for Plant Metabolomics ResearchUniversity of JohannesburgAuckland ParkSouth Africa

Personalised recommendations