Optimizing 1D 1H-NMR profiling of plant samples for high throughput analysis: extract preparation, standardization, automation and spectra processing

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Deborde, Catherine | Fontaine, Jean-Xavier, | Jacob, Daniel | Botana, Adolfo | Nicaise, Valerie | Richard-Forget, Florence | Lecomte, Sylvain | Decourtil, Cédric | Hamade, Kamar | Mesnard, François | Moing, Annick | Molinié, Roland

Edité par HAL CCSD ; Springer Verlag

UMR BFP - Equipe Métabolisme. International audience. Introduction Proton nuclear magnetic resonance spectroscopy (H-1-NMR)-based metabolomic profiling has a range of applications in plant sciences. Objectives The aim of the present work is to provide advice for minimizing uncontrolled variability in plant sample preparation before and during NMR metabolomic profiling, taking into account sample composition, including its specificity in terms of pH and paramagnetic ion concentrations, and NMR spectrometer performances. Methods An automation of spectrometer preparation routine standardization before NMR acquisition campaign was implemented and tested on three plant sample sets (extracts of durum wheat spikelet, Arabidopsis leaf and root, and flax leaf, root and stem). We performed H-1-NMR spectroscopy in three different sites on the wheat sample set utilizing instruments from two manufacturers with different probes and magnetic field strengths. The three collections of spectra were processed separately with the NMRProcFlow web tool using intelligent bucketing, and the resulting buckets were subjected to multivariate analysis. Results Comparability of large- (Arabidopsis) and medium-size (flax) datasets measured at 600MHz and from the wheat sample set recorded at the three sites (400, 500 and 600MHz) was exceptionally good in terms of spectral quality. The coefficient of variation of the full width at half maximum (FWHM) and the signal-to-noise ratio (S/N) of two selected peaks was comprised between 5 and 10% depending on the size of sample set and the spectrometer field. EDTA addition improved citrate and malate resonance patterns for wheat sample sets. A collection of 22 samples of wheat spikelet extracts was used as a proof of concept and showed that the data collected at the three sites on instruments of different field strengths and manufacturers yielded the same discrimination pattern of the biological groups. Conclusion Standardization or automation of several steps from extract preparation to data reduction improves data quality for small to large collections of plant samples of different origins.

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