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First published online September 3, 2004; 10.1104/pp.104.041608

Plant Physiology 136:2641-2651 (2004)
© 2004 American Society of Plant Biologists

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BIOCHEMICAL PROCESSES AND MACROMOLECULAR STRUCTURES

Identification of a Specific Isoform of Tomato Lipoxygenase (TomloxC) Involved in the Generation of Fatty Acid-Derived Flavor Compounds1

Guoping Chen2, Rachel Hackett3, David Walker, Andy Taylor, Zhefeng Lin and Donald Grierson*

Biotechnology and Biological Sciences Research Council Research Group in Plant Gene Regulation, Plant Sciences Division (G.C., R.H., D.W., D.G.), and Food Sciences Division (A.T.), School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicestershire LE12 5RD, United Kingdom

There are at least five lipoxygenases (TomloxA, TomloxB, TomloxC, TomloxD, and TomloxE) present in tomato (Lycopersicon esculentum Mill.) fruit, but their role in generation of fruit flavor volatiles has been unclear. To assess the physiological role of TomloxC in the generation of volatile C6 aldehyde and alcohol flavor compounds, we produced transgenic tomato plants with greatly reduced TomloxC using sense and antisense constructs under control of the cauliflower mosaic virus 35S promoter. The expression level of the TomloxC mRNA in some transgenic plants was selectively reduced by gene silencing or antisense inhibition to between 1% and 5% of the wild-type controls, but the expression levels of mRNAs for the four other isoforms were unaffected. The specific depletion of TomloxC in transgenic tomatoes led to a marked reduction in the levels of known flavor volatiles, including hexanal, hexenal, and hexenol, to as little as 1.5% of those of wild-type controls following maceration of ripening fruit. Addition of linoleic or linolenic acid to fruit homogenates significantly increased the levels of flavor volatiles, but the increase with the TomloxC-depleted transgenic fruit extracts was much lower than with the wild-type control. Confocal imaging of tobacco (Nicotiana tabacum) leaf cells expressing a TomloxC-GFP fusion confirmed a chloroplast localization of the protein. Together, these results suggest that TomloxC is a chloroplast-targeted lipoxygenase isoform that can use both linoleic and linolenic acids as substrates to generate volatile C6 flavor compounds. The roles of the other lipoxygenase isoforms are discussed.

1 This work was supported by the Biotechnology and Biological Science Research Council.

2 Present address: School of Bioengineering, Chongqing University, Chongqing shi, 400044, PR China.

3 Present address: Waltham Centre for Pet Nutrition, Freeby Lane, Melton Mowbray, Leicestershire LE14 4RT, UK.

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.104.041608.

* Corresponding author; e-mail donald.grierson{at}nottingham.ac.uk; fax 0115–9516334.

Received February 25, 2004; returned for revision March 31, 2004; accepted May 28, 2004.




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