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Multifaceted Investigation of Metabolites During Nitrogen Fixation in Medicago via High Resolution MALDI-MS Imaging and ESI-MS

Journal of The American Society for Mass Spectrometry volume 26, pages 149–158 (2015)Cite this article

Abstract

Legumes have developed the unique ability to establish a symbiotic relationship with soil bacteria known as rhizobia. This interaction results in the formation of root nodules in which rhizobia thrive and reduce atmospheric dinitrogen into plant-usable ammonium through biological nitrogen fixation (BNF). Owing to the availability of genetic information for both of the symbiotic partners, the Medicago truncatula–Sinorhizobium meliloti association is an excellent model for examining the BNF process. Although metabolites are important in this symbiotic association, few studies have investigated the array of metabolites that influence this process. Of these studies, most target only a few specific metabolites, the roles of which are either well known or are part of a well-characterized metabolic pathway. Here, we used a multifaceted mass spectrometric (MS) approach to detect and identify the key metabolites that are present during BNF using the Medicago truncatula–Sinorhizobium meliloti association as the model system. High mass accuracy and high resolution matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) Orbitrap instruments were used in this study and provide complementary results for more in-depth characterization of the nitrogen-fixation process. We used well-characterized plant and bacterial mutants to highlight differences between the metabolites that are present in functional versus nonfunctional nodules. Our study highlights the benefits of using a combination of mass spectrometric techniques to detect differences in metabolite composition and the distributions of these metabolites in plant biology.

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Acknowledgments

This work was supported by funding from the University of Wisconsin Graduate School and the Wisconsin Alumni Research Foundation (WARF), a Romnes Faculty Research Fellowship program to L.L. and a National Science Foundation (NSF) grant to J.M.A. (NSF#0701846). E.G. acknowledges an NSF Graduate Research Fellowship (DGE-1256259). The MALDI-Orbitrap and Q-Exactive instruments were purchased through an NIH shared instrument grant (NCRR S10RR029531).

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Authors and Affiliations

  1. Department of Chemistry, University of Wisconsin-Madison, Madison, WI, 53706, USA

    Erin Gemperline & Lingjun Li

  2. Department of Agronomy, University of Wisconsin-Madison, Madison, WI, 53706, USA

    Dhileepkumar Jayaraman, Junko Maeda & Jean-Michel Ané

  3. School of Pharmacy, University of Wisconsin-Madison, Madison, WI, 53705, USA

    Lingjun Li

Authors
  1. Erin Gemperline
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  2. Dhileepkumar Jayaraman
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  3. Junko Maeda
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  4. Jean-Michel Ané
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Correspondence to Lingjun Li.

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Gemperline, E., Jayaraman, D., Maeda, J. et al. Multifaceted Investigation of Metabolites During Nitrogen Fixation in Medicago via High Resolution MALDI-MS Imaging and ESI-MS. J. Am. Soc. Mass Spectrom. 26, 149–158 (2015). https://doi.org/10.1007/s13361-014-1010-0

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  • DOI: https://doi.org/10.1007/s13361-014-1010-0

Keywords

  • Nitrogen fixation
  • Medicago truncatula
  • Metabolites
  • MALDI
  • Orbitrap
  • Mass spectrometry
  • Imaging
  • Q-Exactive