, Volume 231, Issue 6, pp 1495–1504 | Cite as

Metabolic and structural rearrangement during dark-induced autophagy in soybean (Glycine max L.) nodules: an electron microscopy and 31P and 13C nuclear magnetic resonance study

  • Pierre Vauclare
  • Richard Bligny
  • Elisabeth Gout
  • Valentine De Meuron
  • François Widmer
Original Article


The effects of dark-induced stress on the evolution of the soluble metabolites present in senescent soybean (Glycine max L.) nodules were analysed in vitro using 13C- and 31P-NMR spectroscopy. Sucrose and trehalose were the predominant soluble storage carbons. During dark-induced stress, a decline in sugars and some key glycolytic metabolites was observed. Whereas 84% of the sucrose disappeared, only one-half of the trehalose was utilised. This decline coincides with the depletion of Gln, Asn, Ala and with an accumulation of ureides, which reflect a huge reduction of the N2 fixation. Concomitantly, phosphodiesters and compounds like P-choline, a good marker of membrane phospholipids hydrolysis and cell autophagy, accumulated in the nodules. An autophagic process was confirmed by the decrease in cell fatty acid content. In addition, a slight increase in unsaturated fatty acids (oleic and linoleic acids) was observed, probably as a response to peroxidation reactions. Electron microscopy analysis revealed that, despite membranes dismantling, most of the bacteroids seem to be structurally intact. Taken together, our results show that the carbohydrate starvation induced in soybean by dark stress triggers a profound metabolic and structural rearrangement in the infected cells of soybean nodule which is representative of symbiotic cessation.


Peribacteroid membrane Glycine Metabolic NMR Nodules Senescence 



Fructose 6-phosphate


γ-Aminobutyric acid


Glucose 6-phosphate


Glycerol 3-phosphate










Mannose 6-phosphate


Peribacteroid membrane






Scanning electron microscopy


Transmission electron microscopy



We are grateful to Dr. John Lomas (ITODYS UMR 7086, Université Paris-Diderot, Paris 7) for his fine work in correction of English text in the manuscript. We also thank Josiane Bonetti for her excellent bibliographic assistance. This work was supported by grants from the University of Lausanne (Switzerland).

Supplementary material

425_2010_1148_MOESM1_ESM.doc (234 kb)
Supplementary material 1 (DOC 234 kb)
425_2010_1148_MOESM2_ESM.pdf (34 kb)
Supplementary material 2 (PDF 34 kb)


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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Pierre Vauclare
    • 1
    • 3
  • Richard Bligny
    • 2
  • Elisabeth Gout
    • 2
  • Valentine De Meuron
    • 1
  • François Widmer
    • 1
  1. 1.Laboratory of Plant Biology and PhysiologyBiology Building UNILLausanneSwitzerland
  2. 2.Laboratoire de Physiologie Cellulaire Végétale, Unité Mixte de Recherche 5168Institut de Recherche en Technologie et Sciences pour le Vivant CEAGrenoble Cedex 9France
  3. 3.MalissardFrance

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