Plant Cell Advance Online Publication
Published on October 21, 2005; 10.1105/tpc.105.035154
Received June 12, 2005
Returned for revision August 30, 2005
Accepted September 28, 2005
TRANSPARENT TESTA10 Encodes a Laccase-Like Enzyme Involved in Oxidative Polymerization of Flavonoids in Arabidopsis Seed Coat
Lucille Pourcel 1, Jean-Marc Routaboul 1, Lucien Kerhoas 2, Michel Caboche 1, Loïc Lepiniec 1, and Isabelle Debeaujon 1*
1 Laboratoire de Biologie des Semences, Unité Mixte de Recherche 204, Institut National de la Recherche Agronomique/Institut National Agronomique Paris-Grignon, Institut Jean-Pierre Bourgin, 78026 Versailles, France
2 Laboratoire de Phytopharmacie et Médiateurs Chimiques, Institut National de la Recherche Agronomique, 78026 Versailles, France
* To whom correspondence should be addressed. E-mail: debeaujo{at}versailles.inra.fr.
The Arabidopsis thaliana transparent testa10 (tt10) mutant exhibits a delay in developmentally determined browning of the seed coat, also called the testa. Seed coat browning is caused by the oxidation of flavonoids, particularly proanthocyanidins, which are polymers of flavan-3-ol subunits such as epicatechin and catechin. The tt10 mutant seeds accumulate more epicatechin monomers and more soluble proanthocyanidins than wild-type seeds. Moreover, intact testa cells of tt10 cannot trigger H2O2-independent browning in the presence of epicatechin and catechin, in contrast with wild-type cells. UV-visible light detection and mass spectrometry revealed that the major oxidation products obtained with epicatechin alone are yellow dimers called dehydrodiepicatechin A. These products differ from proanthocyanidins in the nature and position of their interflavan linkages. Flavonol composition was also affected in tt10 seeds, which exhibited a higher ratio of quercetin rhamnoside monomers versus dimers than wild-type seeds. We identified the TT10 gene by a candidate gene approach. TT10 encodes a protein with strong similarity to laccase-like polyphenol oxidases. It is expressed essentially in developing testa, where it colocalizes with the flavonoid end products proanthocyanidins and flavonols. Together, these data establish that TT10 is involved in the oxidative polymerization of flavonoids and functions as a laccase-type flavonoid oxidase.
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