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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

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Abstract

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.

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Abbreviations

Fru6P:

Fructose 6-phosphate

GABA:

γ-Aminobutyric acid

Glc6P:

Glucose 6-phosphate

Glyc3P:

Glycerol 3-phosphate

GPC:

Glycerylphosphoryl-choline

GPE:

Glycerylphosphoryl-ethanolamine

GPG:

Glycerylphosphoryl-glycerol

GPI:

Glycerylphosphoryl-inositol

Man6P:

Mannose 6-phosphate

PBM:

Peribacteroid membrane

P-cho:

Phosphoryl-choline

P-eth:

Phosphoryl-ethanolamine

SEM:

Scanning electron microscopy

TEM:

Transmission electron microscopy

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Acknowledgments

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).

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  1. Pierre Vauclare

    Present address: , 22, rue du Lavoir, 26120, Malissard, France

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  1. Laboratory of Plant Biology and Physiology, Biology Building UNIL, room 5449, 1015, Lausanne, Switzerland

    Pierre Vauclare, Valentine De Meuron & François Widmer

  2. Laboratoire de Physiologie Cellulaire Végétale, Unité Mixte de Recherche 5168, Institut de Recherche en Technologie et Sciences pour le Vivant CEA, 17, rue des Martyrs, 38054, Grenoble Cedex 9, France

    Richard Bligny & Elisabeth Gout

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Correspondence to Pierre Vauclare.

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Vauclare, P., Bligny, R., Gout, E. et al. 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. Planta 231, 1495–1504 (2010). https://doi.org/10.1007/s00425-010-1148-3

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