In vivo measurement of the size of oil bodies in plant seeds using a simple and robust pulsed field gradient NMR method

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• 作者：Marina Gromova (1) (2)
  Armel Guillermo (3) (4) (5)
  Pierre-Alain Bayle (1) (2)
  Michel Bardet (1) (2)
  
  1. University of Grenoble Alpes ; INAC-SCIB ; LRM ; 38000 ; Grenoble ; France
  2. CEA ; INAC-SCIB ; LRM ; 38000 ; Grenoble ; France
  3. University of Grenoble Alpes ; INAC-SPrAM ; 38000 ; Grenoble ; France
  4. CNRS ; INAC-SPrAM ; 38000 ; Grenoble ; France
  5. CEA ; INAC-SPrAM ; 38000 ; Grenoble ; France
  
• 关键词：PFGNMR ; Mature seeds ; Oil body size ; In situ determination ; Storage lipids ; Lipid quantification ; TAG
• 刊名：European Biophysics Journal
• 出版年：2015
• 出版时间：April 2015
• 年：2015
• 卷：44
• 期：3
• 页码：121-129
• 全文大小：666 KB
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• 刊物类别：Physics and Astronomy
• 刊物主题：Physics
  Biophysics and Biomedical Physics
  Cell Biology
  Biochemistry
  Plant Physiology
  Animal Physiology
  Neurobiology
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-1017

文摘

An easy to implement and convenient method to measure the mean size of oil bodies (OBs) in plant seeds is proposed using a pulsed field gradient nuclear magnetic resonance (PFGNMR) approach. PFGNMR is a well-known technique used to study either free or restricted diffusion of molecules. As triacylglycerols (TAG) are confined in OBs, analysis of their diffusion properties is a well-suited experimental approach to determine OB sizes. In fact, at long diffusion time, TAG mean squared displacement is limited by the size of the domain where these molecules are confined. In order to access the OB size distribution, strong intensities of magnetic field gradients are generally required. In this work we demonstrate for the first time that a standard liquid-phase NMR probe equipped with a weak-intensity gradient coil can be used to determine the mean size of OBs. Average sizes were measured for several seeds, and OB diameters obtained by PFGNMR were fully consistent with previously published values obtained by microscopy techniques. Moreover, this approach provided evidence of TAG transfer through the network of interconnected OBs, which is dependent on the ability of adjacent membranes to open diffusion routes between OBs. The main advantage of the NMR method is that it does not require any sample preparation and experiments are performed with whole seeds directly introduced in a standard NMR tube.