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Assessment of a 1H high-resolution magic angle spinning NMR spectroscopy procedure for free sugars quantification in intact plant tissue

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Abstract

In most plants, sucrose is the primary product of photosynthesis, the transport form of assimilated carbon, and also one of the main factors determining sweetness in fresh fruits. Traditional methods for sugar quantification (mainly sucrose, glucose and fructose) require obtaining crude plant extracts, which sometimes involve substantial sample manipulation, making the process time-consuming and increasing the risk of sample degradation. Here, we describe and validate a fast method to determine sugar content in intact plant tissue by using high-resolution magic angle spinning nuclear magnetic resonance spectroscopy (HR-MAS NMR). The HR-MAS NMR method was used for quantifying sucrose, glucose and fructose in mesocarp tissues from melon fruits (Cucumis melo var. reticulatus and Cucumis melo var. cantalupensis). The resulting sugar content varied among individual melons, ranging from 1.4 to 7.3 g of sucrose, 0.4–2.5 g of glucose; and 0.73–2.83 g of fructose (values per 100 g fw). These values were in agreement with those described in the literature for melon fruit tissue, and no significant differences were found when comparing them with those obtained using the traditional, enzymatic procedure, on melon tissue extracts. The HR-MAS NMR method offers a fast (usually <30 min) and sensitive method for sugar quantification in intact plant tissues, it requires a small amount of tissue (typically 50 mg fw) and avoids the interferences and risks associated with obtaining plant extracts. Furthermore, this method might also allow the quantification of additional metabolites detectable in the plant tissue NMR spectrum.

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Abbreviations

CoR:

Coefficients of repeatability

CV:

Coefficient of variation

D2O:

Deuterium oxide

FMF:

Focused microwave fixation

fw:

Fresh weight

GC:

Gas chromatography

1H NMR:

Proton NMR

HPLC:

High-performance liquid chromatography

HR-MAS:

High-resolution magic angle spinning

LoA:

Limits of agreement

MS:

Mass spectrometry

NMR:

Nuclear magnetic resonance

qNMR:

Quantitative NMR

SNR:

Signal-to-noise ratio

T1:

Longitudinal relaxation time

TR:

Recycling time

V:

Wilcoxon signed-rank test statistic

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Acknowledgments

We are grateful to Drs. P. Puigdomènech and J. García-Mas for critical reading of this manuscript and to Llorenç Badiella (Servei d’Estadistica, UAB) for initial advice on the statistical analysis of data. This work was funded by grants from the Spanish Ministerio de Ciencia e Innovación (SAF2008-03323 and SAF2011-23870 to CA) and the Ministerio de Economia y Competitividad (BIO2009-08719 and BIO2012-32838 to BSS). CIBER-BBN is an initiative of Instituto de Salud Carlos III, Spain, which is co-funded with EU-funds.

Conflict of interest

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Unitat de Biociències, Dept. Bioquímica i Biologia Molecular, Universitat Autònoma de Barcelona, Edifici C, 08193, Cerdanyola del Vallès, Spain

    Teresa Delgado-Goñi, Juana Martín-Sitjar & Carles Arús

  2. Servei de RMN, Universitat Autònoma de Barcelona, Edifici C, 08193, Cerdanyola del Vallès, Spain

    Miquel E. Cabañas

  3. Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN), Cerdanyola del Vallès, Spain

    Teresa Delgado-Goñi, Juana Martín-Sitjar, Miquel E. Cabañas & Carles Arús

  4. Institut de Biotecnologia i de Biomedicina, Universitat Autònoma de Barcelona, Edifici C, 08193, Cerdanyola del Vallès, Spain

    Carles Arús

  5. Departamento de Genética Molecular, Centre de Recerca en Agrigenòmica (CRAG) CSIC-IRTA-UAB-UB, Edifici CRAG, Campus UAB, Bellaterra, 08193, Cerdanyola del Vallès, Spain

    Sonia Campo & Blanca San Segundo

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  1. Teresa Delgado-Goñi
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  2. Sonia Campo
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  3. Juana Martín-Sitjar
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  4. Miquel E. Cabañas
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  6. Carles Arús
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Correspondence to Carles Arús.

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Delgado-Goñi, T., Campo, S., Martín-Sitjar, J. et al. Assessment of a 1H high-resolution magic angle spinning NMR spectroscopy procedure for free sugars quantification in intact plant tissue. Planta 238, 397–413 (2013). https://doi.org/10.1007/s00425-013-1924-y

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