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First published online October 29, 2004; 10.1104/pp.104.049718

Plant Physiology 136:3504-3514 (2004)
© 2004 American Society of Plant Biologists

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BIOCHEMICAL PROCESSES AND MACROMOLECULAR STRUCTURES

Circadian Regulation of the PhCCD1 Carotenoid Cleavage Dioxygenase Controls Emission of {beta}-Ionone, a Fragrance Volatile of Petunia Flowers1

Andrew J. Simkin, Beverly A. Underwood, Michele Auldridge, Holly M. Loucas, Kenichi Shibuya, Eric Schmelz, David G. Clark and Harry J. Klee*

Horticultural Sciences, Plant Molecular and Cellular Biology Program, University of Florida, Gainesville, Florida 32611–0690 (A.J.S., M.A., H.J.K.); Environmental Horticulture, University of Florida, Gainesville, Florida 32611–0670 (B.A.U., H.M.L., K.S., D.G.C.); and United States Department of Agriculture, Agricultural Research Service, Center for Medical Agricultural and Veterinary Entomology, Gainesville, Florida 32608 (E.S.)

Carotenoids are thought to be the precursors of terpenoid volatile compounds that contribute to flavor and aroma. One such volatile, {beta}-ionone, is important to fragrance in many flowers, including petunia (Petunia hybrida). However, little is known about the factors regulating its synthesis in vivo. The petunia genome contains a gene encoding a 9,10(9',10') carotenoid cleavage dioxygenase, PhCCD1. The PhCCD1 is 94% identical to LeCCD1A, an enzyme responsible for formation of {beta}-ionone in tomato (Lycopersicon esculentum; Simkin AJ, Schwartz SH, Auldridge M, Taylor MG, Klee HJ [2004] Plant J [in press]). Reduction of PhCCD1 transcript levels in transgenic plants led to a 58% to 76% decrease in {beta}-ionone synthesis in the corollas of selected petunia lines, indicating a significant role for this enzyme in volatile synthesis. Quantitative reverse transcription-PCR analysis revealed that PhCCD1 is highly expressed in corollas and leaves, where it constitutes approximately 0.04% and 0.02% of total RNA, respectively. PhCCD1 is light-inducible and exhibits a circadian rhythm in both leaves and flowers. {beta}-Ionone emission by flowers occurred principally during daylight hours, paralleling PhCCD1 expression in corollas. The results indicate that PhCCD1 activity and {beta}-ionone emission are likely regulated at the level of transcript.

1 This work was supported in part by National Science Foundation (grant no. IBN0115004 to H.J.K.).

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.049718.

* Corresponding author; e-mail hjklee{at}ifas.ufl.edu; fax 352–846–2063.

Received July 13, 2004; returned for revision August 20, 2004; accepted August 20, 2004.




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