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. 2011 Aug 16;4(8):1426-1439.
doi: 10.3390/ma4081426.

Low and High Field Magnetic Resonance for in Vivo Analysis of Seeds

Ljudmilla Borisjuk  1 , Hardy Rolletschek  2 , Johannes Fuchs  3   4 , Gerd Melkus  5   6 , Thomas Neuberger  7
Affiliations

Low and High Field Magnetic Resonance for in Vivo Analysis of Seeds

Ljudmilla Borisjuk et al. Materials (Basel). .

Abstract

Low field NMR has been successfully used for the evaluation of seed composition and quality, but largely only in crop species. We show here that 1.5T NMR provides a reliable means for analysing the seed lipid fraction present in a wide range of species, where both the seed size and lipid concentration differed by >10 fold. Little use of high field NMR has been made in seed research to date, even though it potentially offers many opportunities for studying seed development, metabolism and storage. Here we demonstrate how 17.5T and 20T NMR can be applied to image seed structure, and analyse lipid and metabolite distribution. We suggest that further technical developments in NMR/MRI will facilitate significant advances in our understanding of seed biology.

Keywords: 13C; MRI; NMR; crop seed; lipid imaging; seed aging; seed quality; sucrose allocation.

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Figures

Figure 1
Figure 1
Measurement of seed lipid content using the MQ60 device. (a) Correlation of NMR data and conventional assays; (b) Seed/grain investigated by NMR were from Pisum sativum, Trachycarpus fortunei, Glycine max, Robinia pseudoacacia, Brassica napus, Poa pratensis, Dactylorhiza fuchsii, Sinapis alba, Sinapis alba, Avena sativa, Hordeum vulgare.
Figure 2
Figure 2
1H NMR visualization of lipids in naturally aged barley grains stored for three years. (a) Grain used for imaging. Arrows show the placement of the virtual sections (longitudinal 1,2,3 and cross section α, β, γ); Colour-coded visualization of lipid distribution (b) along the grain axis (1,2,3) and (c) across the cross section (α, β, γ).
Figure 3
Figure 3
Quantitative NMR based imaging of lipids in living grains at the start (bottom) and end (top) of the artificial aging procedure. (a, b) Partial 3D representation of lipid distribution in the grain; (c) a lipid map showing the fall in embryo oil content in the aged grain (the red colour indicates the maximum lipid content). a: aleurone, e: endosperm, em: embryo.
Figure 4
Figure 4
Localized 1H NMR and 1H spectroscopic imaging used for the assessment of metabolite distribution within an intact living seed. Reference and metabolite images of the pea seed of DW 120 mg (left) and 250 mg (right). Localized spectra of the pea endosperm (right panel). Upper spectrum: 120 mg DW seed, lower spectrum: 250 mg DW seed. A lower alanine (Ala) and glutamine (Gln) signal was obtained from the younger seed.
Figure 5
Figure 5
L-COSY spectrum of the endosperm of a pea mutant [17] seed.
Figure 6
Figure 6
ge-HMQC spectroscopic imaging of a barley grain. (a) Pulse sequence with water pre-saturation and 13C decoupling; (b) ge-HMQC spectrum from the endosperm region; (c) reference and metabolic images from 13C sucrose after 14h feeding with 13C labelled sucrose.
See this image and copyright information in PMC

References

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