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Subcellular and immunocytochemical localization of the enzymes involved in ammonia assimilation in mesophyll and bundle-sheath cells of maize leaves

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Abstract

The cellular localization of the enzymes involved in primary nitrogen assimilation was investigated following separation of mesophyll protoplasts and bundle-sheath cells of maize (Zea mays L.) leaves. Determination of the enzymatic activities in the two types of cell revealed that nitrate and nitrite reductase are principally located in the mesophyll cells whereas glutamine synthetase (GS) and ferredoxin-dependent glutamate synthase (Fd-GOGAT) are present in both tissues with a preferential location in the bundle-sheath strands. In order to confirm the results obtained by this conventional biochemical method we have used an in-situ immunofluorescence technique to unambiguously localize GS and Fd-GOGAT at the cellular level. Thin-sectioned maize leaves treated with specific GS and Fd-GOGAT antisera followed by conjugation with fluorescein-isothiocyanate-labelled sheep anti-rabbit immunoglobulins clearly show that GS is equally distributed within the leaf whereas Fd-GOGAT is mostly present in the chloroplasts of the bundle-sheath cells. The cellular localization of nitrate reductase, nitrite reductase, GS-2 and Fd-GOGAT in maize leaf cell types strongly indicates that primary nitrogen assimilation functions in the mesophyll cells while photorespiratory nitrogen recycling is restricted to the bundle-sheath cells.

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Abbreviations

BSC:

bundle-sheath cells

Chl:

chlorophyll

Fd-GOGAT:

ferredoxin-dependent glutamate synthase

FITC:

fluorescein isothiocyanate

GS-1, GS-2:

cytosolic and chloroplas-tic glutamine synthetase, respectively

NR:

nitrate reductase

MC:

mesophyll cells

NiR:

itrite reductase

Rubisco:

ribulose-1,5-bisphosphate carboxylase oxygenase

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Authors and Affiliations

  1. Laboratoire de Biologie Cellulaire, INRA, Centre de Versailles, Route de Saint-Cyr, F-78026, Versailles Cedex, France

    Thomas W. Becker

  2. Institut des Sciences Végétales, Avenue de la Terrasse, F-91190, Gif sur Yvette Cedex, France

    Catherine Perrot-Rechenmann

  3. Laboratoire du Métabolisme et de la Nutrition des Plantes INRA, Centre de Versailles, Route de Saint-Cyr, F-78026, Versailles Cedex, France

    Akira Suzuki & Bertrand Hirel

Authors
  1. Thomas W. Becker
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  2. Catherine Perrot-Rechenmann
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  3. Akira Suzuki
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  4. Bertrand Hirel
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We wish to thank Dr. Christine Foyer and Marie-Claude Marsolier, (Laboratoire du Métabolisme et de la Nutrition des Plantes, INRA, Versailles, France), for critical reading of the manuscript. T.W. Becker was supported by a Biotechnology Action Program research grant from the Commission of the European Communities.

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Becker, T.W., Perrot-Rechenmann, C., Suzuki, A. et al. Subcellular and immunocytochemical localization of the enzymes involved in ammonia assimilation in mesophyll and bundle-sheath cells of maize leaves. Planta 191, 129–136 (1993). https://doi.org/10.1007/BF00240904

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