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1H NMR and GC-MS metabolic fingerprinting of developmental stages of Rhizoctonia solani sclerotia

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Abstract

Rhizoctonia solani AG-3 is a soilborne plant pathogen that forms resting vegetative structures called sclerotia. These compact structures are crucial to the pathogen’s survival and pathogenesis. The metabolic changes occurring during sclerotia development were monitored using proton nuclear magnetic resonance (1H NMR) spectroscopy and gas chromatography–mass spectrometry (GC-MS). The validation, discrimination, and the establishment of correlative relationships between metabolite signals were performed by principal component analysis (PCA) and partial least squares-discriminant analysis (PLS-DA). The results of the analyses suggested that out of the 116 compounds that were simultaneously analyzed and compared using GC-MS, α-α-trehalose, d-glucose, 9-(Z)-octadecenoic and 9,12-octadecadienoic acids, xylitol, and glucitol were key metabolites that were highly dependent on the developmental stage of the sclerotia contributing to their discrimination and classification. Furthermore, the application of 1H NMR and GC-MS metabolic fingerprinting on the same biological sample provided complementary information illustrating the value of this integrated approach in the study of metabolic changes in fungal structures.

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Acknowledgments

The authors thank Dr. W.D. Marshall for providing access to GC-MS instrument and Dr. T. Sprules, the Quebec/Eastern Canada High Field NMR Facility, for her assistance with the NMR spectra. Funding for this was provided by the Natural Sciences and Engineering Research Council of Canada (NSERC) discovery Grants and a Post-doctoral fellowship to Dr. K. Aliferis funded by the Fonds Québécois de la Recherche sur la Nature et les Technologies (FQRNT) and McGill University.

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  1. Department of Plant Science, McGill University, 21111 Lakeshore Road, Sainte-Anne-de-Bellevue, QC, H9X 3V9, Canada

    K. A. Aliferis & S. Jabaji

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  1. K. A. Aliferis
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  2. S. Jabaji
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Correspondence to S. Jabaji.

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11306_2009_180_MOESM1_ESM.jpg

Predicted 1H NMR spectra of representative compounds identified by GC-MS in sclerotial extracts of Rhizoctonia solani using the ACD/C+H NMR Predictor and Database v.12.01 (ACD/Labs) (JPG 1,568 kb)

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Aliferis, K.A., Jabaji, S. 1H NMR and GC-MS metabolic fingerprinting of developmental stages of Rhizoctonia solani sclerotia. Metabolomics 6, 96–108 (2010). https://doi.org/10.1007/s11306-009-0180-4

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