Plant Molecular Biology

, Volume 31, Issue 4, pp 803–817 | Cite as

Localization of tobacco cytosolic glutamine synthetase enzymes and the corresponding transcripts shows organ- and cell-specific patterns of protein synthesis and gene expression

  • Frédéric Dubois
  • Norbert Brugière
  • Rajbir S. Sangwan
  • Bertrand Hirel
Article

Abstract

The subcellular localization of glutamine synthetase in tobacco and the differential expression of two genes encoding cytosolic enzyme was investigated using both immunocytochemistry and in situ hybridization. Two full length cDNA clones each encoding cytosolic GS (Glnl-3 and Glnl-5) were isolated from a tobacco seedling cDNA library. A strong homology was found in the coding region of the two clones whereas the 3′- and 5′-untranslated sequences were dissimilar. In order to determine the levels of transcription, specific sequences from Glnl-3 and Glnl-5 were used in an RNAse protection assay. This experiment clearly showed that the gene encoding Glnl-3 is expressed in roots and flowers whereas the gene encoding Glnl-5 is transcribed at a high level in stems and at a lower level in roots and flowers. Immunogold labelling was used to examine the subcellular and cellular distribution of glutamine synthetase in vegetative and reproductive organs of tobacco plants. In mature leaf tissue or petals and sepals, plastidic GS was visualised only in the stroma matrix of chloroplasts and plastids. Cytosolic GS was detected in a number of vegetative or reproductive organs including leaves and flowers. In leaves cytosolic GS was preferentially located in the vascular tissue. In situ hybridization was performed using sections of tobacco organs and specific antisense RNA probes to the genes encoding Glnl-3 and Glnl-5. Glnl-5 transcripts were localised in the vascular tissues of stems and roots whereas Glnl-3 transcripts were detected in all root cells and floral organs including petals, sepals and anthers.

Key words

glutamine synthetase immunocytochemistry flowers in situ hybridization Nicotiana tabacum L. phloem 

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Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Frédéric Dubois
    • 1
  • Norbert Brugière
    • 2
  • Rajbir S. Sangwan
    • 1
  • Bertrand Hirel
    • 2
  1. 1.Laboratoire d'Androgenèse et BiotechnologieUniversité de Picardie Jules VerneAmiens CedexFrance
  2. 2.Laboratoire du Métabolisme et de la Nutrition des Plantes, INRACentre de VersaillesVersailles CedexFrance

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