First published online December 23, 2005; 10.1104/pp.105.073262
Plant Physiology 140:279-291 (2006)
© 2006 American Society of Plant Biologists
SYSTEMS BIOLOGY, MOLECULAR BIOLOGY, AND GENE REGULATION
Identification of the Flavonoid Hydroxylases from Grapevine and Their Regulation during Fruit Development1
Jochen Bogs,
Ali Ebadi2,
Debra McDavid and
Simon P. Robinson*
Commonwealth Scientific and Industrial Research Organization, Plant Industry, Horticulture Unit, Glen Osmond, South Australia 5064, Australia (J.B., A.E., D.M., S.P.R.); and Cooperative Research Centre for Viticulture, Glen Osmond, South Australia 5064, Australia (J.B., D.M., S.P.R.)
Flavonoids are important secondary metabolites in many fruits, and their hydroxylation pattern determines their color, stability, and antioxidant capacity. Hydroxylation of the B-ring of flavonoids is catalyzed by flavonoid 3'-hydroxylase (F3'H) and flavonoid 3',5'-hydroxylase (F3'5'H), and may also require cytochrome b5. We report the identification of genes encoding F3'H, F3'5'H, and a putative cytochrome b5 from grapevine (Vitis vinifera L. cv Shiraz) and their transcriptional regulation in fruit. Functionality of the genes VvF3'H and VvF3'5'H1 was demonstrated by ectopic expression in petunia (Petunia hybrida), which altered flower color and flavonoid composition as expected. VvF3'H was expressed in grapes before flowering, when 3'-hydroxylated flavonols are made, and all three genes were expressed after flowering, when proanthocyanidins (PAs) are synthesized. In berry skin, expression of all three genes was low at the onset of ripening (véraison) but increased after véraison concomitant with the accumulation of 3'- and 3',5'-hydroxylated anthocyanins. VvF3'H and VvCytoB5 were expressed in seeds but not VvF3'5'H1, consistent with the accumulation of 3'-hydroxylated PAs in this tissue. VvCytoB5 expression was correlated with expression of both VvF3'H and VvF3'5'H1 in the different grape tissues. In contrast to red grapes, where VvF3'H, VvF3'5'H1, and VvCytoB5 were highly expressed during ripening, the expression of VvF3'5'H1 and VvCytoB5 in white grapes during ripening was extremely low, suggesting a difference in transcriptional regulation. Our results show that temporal and tissue-specific expression of VvF3'H, VvF3'5'H1, and VvCytoB5 in grapes is coordinated with the accumulation of the respective hydroxylated flavonols and PAs, as well as anthocyanins. Understanding the regulation of flavonoid hydroxylases could be used to modify flavonoid composition of fruits.
1 This work was supported by the Australian Government's Cooperative Research Centres Program and the Grape and Wine Research and Development Corporation.
2 Present address: Department of Horticulture, Faculty of Agriculture, University of Tehran, Karaj, Iran.
The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Simon P. Robinson (simon.robinson{at}csiro.au).
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.105.073262.
* Corresponding author; e-mail simon.robinson{at}csiro.au; fax 61–8–8303–8601.
Received October 24, 2005;
returned for revision October 24, 2005;
accepted November 10, 2005.
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