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Planta

, Volume 238, Issue 2, pp 397–413 | Cite as

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

  • Teresa Delgado-Goñi
  • Sonia Campo
  • Juana Martín-Sitjar
  • Miquel E. Cabañas
  • Blanca San Segundo
  • Carles Arús
Emerging Technologies

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.

Keywords

HR-MAS NMR Sugar quantification Melon mesocarp Enzymatic analysis Differences vs averages method 

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

Notes

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.

Supplementary material

425_2013_1924_MOESM1_ESM.pdf (1.8 mb)
Supplementary material 1 (PDF 1810 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Teresa Delgado-Goñi
    • 1
    • 3
  • Sonia Campo
    • 5
  • Juana Martín-Sitjar
    • 1
    • 3
  • Miquel E. Cabañas
    • 2
    • 3
  • Blanca San Segundo
    • 5
  • Carles Arús
    • 1
    • 3
    • 4
  1. 1.Unitat de Biociències, Dept. Bioquímica i Biologia MolecularUniversitat Autònoma de BarcelonaCerdanyola del VallèsSpain
  2. 2.Servei de RMNUniversitat Autònoma de BarcelonaCerdanyola del VallèsSpain
  3. 3.Centro de Investigación Biomédica en Red en Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN)Cerdanyola del VallèsSpain
  4. 4.Institut de Biotecnologia i de BiomedicinaUniversitat Autònoma de BarcelonaCerdanyola del VallèsSpain
  5. 5.Departamento de Genética MolecularCentre de Recerca en Agrigenòmica (CRAG) CSIC-IRTA-UAB-UBCerdanyola del VallèsSpain

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