TRANSPARENT TESTA10 Encodes a Laccase-Like Enzyme Involved in Oxidative Polymerization of Flavonoids in Arabidopsis Seed Coat

• Published in: The Plant cell Vol. 17; no. 11; pp. 2966 - 2980
• Main Authors: Pourcel, Lucille, Routaboul, Jean-Marc, Kerhoas, Lucien, Caboche, Michel, Lepiniec, Loi̊c, Debeaujon, Isabelle
• Format: Journal Article
• Language: English
• Published: United States American Society of Plant Biologists 01.11.2005; American Society of Plant Biologists (ASPB)
• Subjects: Alleles; Amino Acid Sequence; amino acid sequences; Arabidopsis - enzymology; Arabidopsis - genetics; Arabidopsis - growth & development; Arabidopsis Proteins - genetics; Arabidopsis Proteins - isolation & purification; Arabidopsis Proteins - metabolism; Arabidopsis thaliana; Base Sequence; Catechol Oxidase - genetics; Catechol Oxidase - isolation & purification; Catechol Oxidase - metabolism; complementary DNA; DNA, Complementary - analysis; DNA, Complementary - genetics; enzyme activity; Enzymes; exons; Flavonoids; Flavonoids - metabolism; Flavonols; gene expression; Gene Expression Regulation, Enzymologic - physiology; Gene Expression Regulation, Plant - physiology; genes; Genetics; introns; laccase; Laccase - genetics; Laccase - isolation & purification; Laccase - metabolism; Life Sciences; messenger RNA; Molecular Sequence Data; mutants; Mutation - genetics; nucleotide sequences; Oxidases; Oxidation; Oxidation-Reduction; oxidoreductases; Oxidoreductases - genetics; Oxidoreductases - isolation & purification; Oxidoreductases - metabolism; Plants; Plants genetics; Polymerization; Polymers - metabolism; Polyphenols; promoter regions; Promoter Regions, Genetic - genetics; Seeds - enzymology; Seeds - genetics; Testa; TT10 gene; POLIMERISATION; FLAVONOID; PROANTHOCYANIDINS; ARABIDOPSIS; TRANSPARENT TESTA 10
• ISSN: 1040-4651; 1532-298X; 1532-298X
• DOI: 10.1105/tpc.105.035154

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.