First published online August 6, 2004; 10.1104/pp.104.041947
Plant Physiology 135:2241-2260 (2004)
© 2004 American Society of Plant Biologists
DEVELOPMENT AND HORMONE ACTION
Proteome Reference Maps of Vegetative Tissues in Pea. An Investigation of Nitrogen Mobilization from Leaves during Seed Filling1
Séverine Schiltz*,
Karine Gallardo,
Myriam Huart,
Luc Negroni,
Nicolas Sommerer and
Judith Burstin
Unité de Génétique et Ecophysiologie des Légumineuses à Graines, Institut National de la Recherche Agronomique, 21065 Dijon cedex, France (S.S., K.G., M.H., J.B.); Proteomic Platform, Unité Mixte de Recherche 320, Institut National de la Recherche Agronomique–Centre National de la Recherche Scientifique–Institut National Agronomique Paris–Grignon, Ferme du Moulon 91190 Gif sur Yvette, France (L.N.); and Unité Mixte de Recherche 5004, Agro–Montpellier–Centre National de la Recherche Scientifique–Institut National de la Recherche Agronomique–Université Montpellier II, 34060 Montpellier cedex 1, France (N.S.)
A proteomic approach was used to analyze protein changes during nitrogen mobilization (N mobilization) from leaves to filling seeds in pea (Pisum sativum). First, proteome reference maps were established for mature leaves and stems. They displayed around 190 Coomassie Blue-stained spots with pIs from 4 to 7. A total of 130 spots were identified by mass spectrometry as corresponding to 80 different proteins implicated in a variety of cellular functions. Although the leaf proteome map contained more abundant spots, corresponding to proteins involved in energy/carbon metabolism, than the stem map, their comparison revealed a highly similar protein profile. Second, the leaf proteome map was used to analyze quantitative variations in leaf proteins during N mobilization. Forty percent of the spots showed significant changes in their relative abundance in the total protein extract. The results confirmed the importance of Rubisco as a source of mobilizable nitrogen, and suggested that in pea leaves the rate of degradation of Rubisco may vary throughout N mobilization. Correlated with the loss of Rubisco was an increase in relative abundance of chloroplastic protease regulatory subunits. Concomitantly, the relative abundance of some proteins related to the photosynthetic apparatus (Rubisco activase, Rubisco-binding proteins) and of several chaperones increased. A role for these proteins in the maintenance of a Rubisco activation state and in the PSII repair during the intense proteolytic activity within the chloroplasts was proposed. Finally, two 14-3-3-like proteins, with a potential regulatory role, displayed differential expression patterns during the massive remobilization of nitrogen.
1 This work was supported in part by the Regional Council of Burgundy, France
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.041947.
* Corresponding author; e-mail schiltz{at}epoisses.inra.fr; fax 33–3–80–693222.
Received March 2, 2004;
returned for revision May 27, 2004;
accepted June 11, 2004.
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