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Published on June 13, 2008; 10.1104/pp.108.116905

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Received January 26, 2008
Accepted June 10, 2008

A Proteomic Profiling Approach to Reveal a Novel Role of BnD22 (Brassica napus drought 22 kD)/Water Soluble Chlorophyll binding Protein in young leaves during Nitrogen Remobilization induced by Stressful conditions

Marie DESCLOS , Lucie DUBOUSSET , Philippe ETIENNE , Francoise LE CAHEREC , Hiroyushi SATOH , Josette BONNEFOY , Alain OURRY , and Jean-Christophe AVICE *

UMR INRA/UCBN 950 Ecophysiologie Vegetale, Agronomie (EVA) et nutritions N C S, IFR 146 ICORE, Institut de Biologie Fondamentale et Appliquee, Universite de CAEN Basse-Normandie, F-14032 Caen, France; UMR INRA/Agrocampus Rennes/Universite de Rennes 1 118 Amelioration des Plantes et Biotechnologies (APBV), Campus de Beaulieu, avenue du General Leclerc, 35042 Rennes cedex, France; Department of Biomolecular Science, Toho University, 2-2-1 Miyama, Funabashi, Chiba 274-8510, Japan

* Corresponding author; email: jean-christophe.avice{at}unicaen.fr.

Despite its Water Soluble Chlorophyll binding Protein (WSCP) function, the putative trypsin inhibitor (TI) activity of the BnD22 (Brassica napus Drought 22 kD) protein and its physiological function in young leaves during leaf N remobilization promoted by stressful conditions remains an enigma. Therefore, our objectives were to determine (i) if BnD22 is related to the 19 kD-TI previously detected in Brassica napus young leaves, and (ii) if the levels of BnD22 transcripts, BnD22 protein and TI activity in young leaves are associated with plant responses to stress conditions (N starvation and MeJA treatments) that are able to modulate leaf senescence. Compared to control, N starvation delayed initiation of senescence and induced 19 kD-TI activity in the young leaves. After 3 d with MeJA, the 19 kD-TI activity was 7-fold higher than the control. Using 2-DE gel, TI activity and ESI-LC MS/MS analysis, it was demonstrated that two 19 kD proteins with pI 5.0 and 5.1 harbouring TI activity, correspond BnD22 perfectly. BnD22 gene expression, TI activities and BnD22 protein presented similar patterns. Using polyclonal anti-WSCP antibodies of Brassica oleracea, six polypeptides separated by 2-DE were detected in young leaves treated with MeJA. ESI-LC MS/MS analysis of six polypeptides confirms their homologies with WSCP. Results suggest that BnD22 possesses dual functions (WSCP and TI) that lead to protect younger tissues from adverse conditions by maintaining metabolism (protein integrity and photosynthesis). By sustaining sink growth of stressed plants, BnD22 may contribute to a better utilization of recycling N from sources, a physiological trait that improves the N use efficiency.






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