Comprehensive analysis of organic ligands in whole root exudates using nuclear magnetic resonance and gas chromatography - Mass spectrometry

Teresa W.M. Fan, Andrew N. Lane, Judy Pedler, David Crowley, Richard M. Higashi

Research output: Contribution to journalArticlepeer-review

122 Scopus citations


Root exudates in the rhizosphere are vital to the normal life cycle of plants. A key factor is phytometallophores, which function in the nutritional acquisition of iron and zinc and are likely to be important in the uptake of pollutant metals by plants. Unraveling the biochemistry of these compounds is tedious using traditional analyses, which also fall short in providing the overall chemical composition or in detecting unknown or unexpected organic ligands in the exudates. Here, we demonstrate a comprehensive analysis of the exudate composition directly by 1H and 13C multidimensional NMR and silylation GC-MS. The advantages are (a) minimal sample preparation, with no loss of unknown compounds, and reduced net analysis time; (b) structure- based analysis for universal detection and identification; and (c) simultaneous analysis of a large number of constituents in a complex mixture. Using barley root exudates, a large number of common organic and amino acids were identified. Three derivatives of mugineic acid phytosiderophores were also determined, the major one being 3-epihydroxymugineic acid, for which complete 1H and 13C NMR assignments were obtained. Quantification of all major components using these methods revealed a sevenfold increase in total exudation under moderate iron deficiency, with 3-epihydroxymugineic acid comprising approximately 22% of the exudate mixture. As iron deficiency increased, total quantities of exudate per gram of root remained unchanged, but the relative quantity of carbon allocated to phytosiderophore increased to approximately 50% of the total exudate in response to severe iron deficiency.

Original languageEnglish
Pages (from-to)57-68
Number of pages12
JournalAnalytical Biochemistry
Issue number1
StatePublished - Aug 15 1997

Bibliographical note

Funding Information:
This work was supported, in part, by U.S. Department of Energy Grant DE-FG07-96ER20255 and the U.C. Davis Center for Ecological Health Research (U.S. Environmental Protection Agency Grant R819658). We acknowledge the U.K. Medical Research Council Biomedical NMR Centre for NMR instrumentation and Dr. M. J. Grad-well for assistance with the HMBC experiments.

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology


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