Plant Molecular Biology

, Volume 62, Issue 1–2, pp 53–69 | Cite as

Spatial and Temporal Organization of Sucrose Metabolism in Lotus japonicus Nitrogen-Fixing Nodules Suggests a Role for the Elusive Alkaline/Neutral Invertase

  • Emmanouil Flemetakis
  • Rodica C. Efrose
  • Thomas Ott
  • Catalina Stedel
  • Georgios Aivalakis
  • Michael K. Udvardi
  • Panagiotis Katinakis
Article

Abstract

Symbiotic nitrogen fixation (SNF) in legume nodules is a highly energy demanding process, fuelled by plant-supplied carbohydrates mainly in the form of sucrose. In this study, we have combined molecular and biochemical approaches in order to study the spatial and temporal organisation of sucrose metabolism in nitrogen-fixing nodules of the model legume Lotus japonicus, with an emphasis on the neglected role of alkaline/neutral invertase. For this purpose, a full-length cDNA clone coding for an alkaline/neutral invertase isoform, termed LjInv1, was identified in a L. japonicus mature nodule cDNA libraries. Alkaline/neutral invertase activity was also found to be the predominant invertase activity in mature nodules. Real-time reverse-transcription polymerase chain reaction analysis was used in order to study the temporal expression patterns of LjInv1 in parallel with genes encoding acid invertase and sucrose synthase (SuSy) isoforms, and enzymes involved in the subsequent hexose partitioning including hexokinase, phosphoglucomutase (PGM) and phosphoglucose isomerase (PGI). The spatial organisation of sucrose metabolism was studied by in situ localisation of LjInv1 transcripts and alkaline/neutral invertase activity, and SuSy protein during nodule development. Furthermore, the spatial organisation of hexose metabolism was investigated by histochemical localisation of hexokinase, PGM and PGI activities in mature nodules. The results considered together indicate that alkaline/neutral invertase could contribute to both the Glc-1-P and Glc-6-P pools in nodules, fuelling both biosynthetic processes and SNF. Furthermore, transcript profiling analysis revealed that genes coding for hexokinase and putative plastidic PGM and PGI isoforms are upregulated during the early stages of nodule development, while the levels of transcripts corresponding to cytosolic PGM and PGI isoforms remained similar to uninfected roots, indicating a possible role of LjInv1 in producing hexoses for starch production and other biosynthetic processes in developing nodules.

Key words

Symbiotic nitrogen fixation Plant–microbe interactions Sucrose metabolism Nodule Symbiosis 

Abbreviations

Fru

fructose

Glc

glucose

Glc-1-P

Glucose-1-phosphate

Glc-6-P

Glucose-6-phosphate

Inv

invertase

PGI

phosphoglucose isomerase

PGM

phosphoglucomutase

SNF

symbiotic nitrogen fixation

Suc

Sucrose

SuSy

sucrose synthase

Notes

Acknowledgements

This work was supported by EU RTN programme MRTN-CT-2003-505227.

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Copyright information

© Springer Science+Business Media B.V. 2006

Authors and Affiliations

  • Emmanouil Flemetakis
    • 1
  • Rodica C. Efrose
    • 1
  • Thomas Ott
    • 2
  • Catalina Stedel
    • 1
  • Georgios Aivalakis
    • 1
  • Michael K. Udvardi
    • 2
  • Panagiotis Katinakis
    • 1
  1. 1.Department of Agricultural BiotechnologyAgricultural University of AthensAthensGreece
  2. 2.Max-Planck-Institute for Molecular Plant PhysiologyGolmGermany

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