Abstract
Methoxypyrazines (MPs) are volatile, grape-derived aroma compounds that contribute to the distinct herbaceous characters of some wines. Although the full pathway leading to MP production has not been elucidated, there is strong evidence that the final step involves the methylation of non-volatile hydroxypyrazine (HP) precursors. Two cDNA encoding O-methyltransferases (OMTs) that have homology to an enzyme previously purified and shown to catalyse the methylation of HPs were isolated from Cabernet Sauvignon. Recombinant protein from the cDNAs (VvOMT1 and VvOMT2) was produced in E. coli and activity assays demonstrated that both encode OMTs able to methylate HPs to produce MPs, however both showed greatest activity against the flavonol quercetin. VvOMT1 has higher catalytic activity against isobutyl hydroxypyrazine compared to isopropyl hydroxypyrazine, whereas the converse is true for VvOMT2. The timing of the expression of VvOMT1 in the skin and the flesh of developing Cabernet Sauvignon grape berries was associated with the period of MP accumulation in these tissues, while VvOMT2 expression was greatest in roots, which were found to contain high levels of MPs. The MP composition of these tissues also reflects the relative levels of expression of these genes and their substrate preference. The identification of genes responsible for MP production in grapevine will help in understanding the effect of different viticultural and environmental factors on MP accumulation.
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Acknowledgments
We wish to thank Dr. Sally Duck of the Commercial Analytical Services at Monash University School of Chemistry, for acquiring high-resolution electrospray-ionization mass spectrometry of HP substrates. We sincerely thank Sue Maffei and Emily Nicholson for their expertise and assistance with GC–MS analysis and Maria Mrinak for her tireless laboratory assistance. We are grateful to the Harvey family of Chalk Hill Wines and to Yalumba for the use of their vineyards. We acknowledge the Grape and Wine Research and Development Corporation for their financial support of this work.
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An erratum to this article can be found online at http://dx.doi.org/10.1007/s11103-013-0012-9.
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Dunlevy, J.D., Soole, K.L., Perkins, M.V. et al. Two O-methyltransferases involved in the biosynthesis of methoxypyrazines: grape-derived aroma compounds important to wine flavour. Plant Mol Biol 74, 77–89 (2010). https://doi.org/10.1007/s11103-010-9655-y
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DOI: https://doi.org/10.1007/s11103-010-9655-y