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First published online October 10, 2003; 10.1105/tpc.014043

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The Plant Cell, Vol. 15, 2514-2531, November 2003, www.plantcell.org ©2003, American Society of Plant Biologists

Proanthocyanidin-Accumulating Cells in Arabidopsis Testa: Regulation of Differentiation and Role in Seed Development

Isabelle Debeaujona, Nathalie Nesi1,a, Pascual Perezb, Martine Devicc, Olivier Grandjeand, Michel Cabochea and Loïc Lepiniec2,a

a Laboratoire de Biologie des Semences, Unité Mixte de Recherche 204 Institut National de la Recherche Agronomique/Institut National Agronomique Paris-Grignon, 78026 Versailles, France
b Laboratoire de Biologie Cellulaire et Moléculaire, Biogemma, 63170 Aubière, France
c Laboratoire Génome et Développement des Plantes, Unité Mixte de Recherche 5096 Centre National de la Recherche Scientifique/Université Perpignan, 66860 Perpignan, France
d Laboratoire de Biologie Cellulaire, Institut National de la Recherche Agronomique, 78026 Versailles, France

2 To whom correspondence should be addressed. E-mail lepiniec{at}versailles.inra.fr; fax 33-1-30-83-30-99

Anthocyanidin reductase encoded by the BANYULS (BAN) gene is the core enzyme in proanthocyanidin (PA) biosynthesis. Here, we analyzed the developmental mechanisms that regulate the spatiotemporal expression of BAN in the developing Arabidopsis seed coat. PA-accumulating cells were localized histochemically in the inner integument (seed body and micropyle) and pigment strand (chalaza). BAN promoter activity was detected specifically in these cells. Gain-of-function experiments showed that an 86-bp promoter fragment functioned as an enhancer specific for PA-accumulating cells. Mutations in regulatory genes of PA biosynthesis abolished BAN promoter activity (transparent testa2 [tt2], tt8, and transparent testa glabra1 [ttg1]), modified its spatial pattern (tt1 and tt16), or had no influence (ttg2), thus revealing complex regulatory interactions at several developmental levels. Genetic ablation of PA-accumulating cells targeted by the BAN promoter fused to BARNASE led to the formation of normal plants that produced viable yellow seeds. Importantly, these seeds had no obvious defects in endosperm and embryo development.




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