Following plant metabolism in vivo and in extracts with heteronuclear two-dimensional nuclear magnetic resonance spectroscopy

Analytical Biochemistry
Y Shachar-HillM W Germann

Abstract

Limits of sensitivity and spectral resolution currently restrict the application of nuclear magnetic resonance (NMR) spectroscopy in plant metabolism. This study shows that these limits can be substantially expanded through the application of heteronuclear single- and multiple-quantum two-dimensional (2D) spectroscopic methods using pulsed field gradients both in vivo and in extracts. The course of metabolism in approximately 0.2 g of maize (Zea mays L.) root tips labeled with [1-13C]glucose was followed with 1 min time resolution using heteronuclear multiple quantum coherence (HMQC) 13C-1H spectroscopy in vivo. The timing of alanine, lactate, and ethanol synthesis was followed during the transition from normal to hypoxic conditions. In extracts of labeled maize root tips, 13C-1H heteronuclear single quantum coherence and heteronuclear multiple quantum coherence (HMBC) spectra acquired in 2-3 h allowed the detection and assignment of resonance that are not seen in one-dimensional (1D) 13C NMR spectra of the same samples taken in 12 h. In root tips labeled with 15NH4+, 15N-(1)H HMQC spectra in vivo showed labeling in the amide of glutamine. In extracts, 15N labeling in amines and amides was detected using 15N-1H HMBC spectra tha...Continue Reading

Citations

May 5, 2001·Annual Review of Plant Physiology and Plant Molecular Biology·R George Ratcliffe, Yair Shachar-Hill
Feb 13, 2016·Plant & Cell Physiology·Satomi KannoLaurent Nussaume
May 1, 2007·Phytochemistry·Stéphane MassouJean-Charles Portais
Feb 25, 2005·Biological Reviews of the Cambridge Philosophical Society·R G Ratcliffe, Y Shachar-Hill
Sep 7, 2005·Photosynthesis Research·F Mesnard, R G Ratcliffe
Aug 10, 2000·Neoplasia : an International Journal for Oncology Research·J KurhanewiczS J Nelson
May 29, 2007·Phytochemistry·Doug K AllenJohn B Ohlrogge
Jul 18, 2001·Journal of the American Chemical Society·C HinseJ H Stöckigt

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