First published online July 30, 2004; 10.1104/pp.104.045468
Plant Physiology 135:1993-2011 (2004)
© 2004 American Society of Plant Biologists
Understanding in Vivo Benzenoid Metabolism in Petunia Petal Tissue1
Jennifer Boatright,
Florence Negre,
Xinlu Chen,
Christine M. Kish,
Barbara Wood,
Greg Peel,
Irina Orlova,
David Gang,
David Rhodes and
Natalia Dudareva*
Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, Indiana 47907 (J.B., F.N., X.C., C.M.K., B.W., G.P., I.O., D.R., N.D.); and Department of Plant Sciences and Institute for Biomedical Science and Biotechnology, University of Arizona, Tucson, Arizona 85721 (D.G.)
In vivo stable isotope labeling and computer-assisted metabolic flux analysis were used to investigate the metabolic pathways in petunia (Petunia hybrida) cv Mitchell leading from Phe to benzenoid compounds, a process that requires the shortening of the side chain by a C2 unit. Deuterium-labeled Phe (2H5-Phe) was supplied to excised petunia petals. The intracellular pools of benzenoid/phenylpropanoid-related compounds (intermediates and end products) as well as volatile end products within the floral bouquet were analyzed for pool sizes and labeling kinetics by gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry. Modeling of the benzenoid network revealed that both the CoA-dependent,  -oxidative and CoA-independent, non--oxidative pathways contribute to the formation of benzenoid compounds in petunia flowers. The flux through the CoA-independent, non--oxidative pathway with benzaldehyde as a key intermediate was estimated to be about 2 times higher than the flux through the CoA-dependent, -oxidative pathway. Modeling of 2H5-Phe labeling data predicted that in addition to benzaldehyde, benzylbenzoate is an intermediate between L-Phe and benzoic acid. Benzylbenzoate is the result of benzoylation of benzyl alcohol, for which activity was detected in petunia petals. A cDNA encoding a benzoyl-CoA:benzyl alcohol/phenylethanol benzoyltransferase was isolated from petunia cv Mitchell using a functional genomic approach. Biochemical characterization of a purified recombinant benzoyl-CoA:benzyl alcohol/phenylethanol benzoyltransferase protein showed that it can produce benzylbenzoate and phenylethyl benzoate, both present in petunia corollas, with similar catalytic efficiencies.
1 This work was supported by the U.S. Department of Agriculture National Research Initiative (grant no. 2003–35318–13619) and by the Fred Gloeckner Foundation (grant to N.D.). This paper is contribution Number 17397 from the Purdue University Agricultural Experimental Station.
Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.045468.
* Corresponding author; e-mail dudareva{at}hort.purdue.edu; fax 765–494–0391.
Received April 28, 2004;
returned for revision June 23, 2004;
accepted June 24, 2004.
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