The aim of this study was to discover genes involved in grape flavour pathways by elicitation of grape cell suspension. Among numerous elicitors tested, methyl jasmonate (MeJA) and jasmonic acid (JA) induce a wide range of sesquiterpenes in ‘Cabernet Sauvignon’ and Riesling berry cell suspension cultures. Sesquiterpenes, the largest subclass of natural terpenes compounds, may be synthesised by two different pathways: the mevalonate pathways (MVA) localised in the cytosol and the methylerytritol phoshate pathway (DOXP/MEP) localised in the plastids. Using specific inhibitors we found that in our cell suspension MeJA induced sequiterpenes are synthesised by the MVA pathway. A set of experiments performed to optimize sesquiterpenes production, indicated that the efficacy of MeJA concentration is strictly dependent upon the packed cell volume (PCV) of cell suspension at the moment of elicitor addition. In time course studies it was found that the peak in sesquiterpene accumulation was recorded 72 hours after MeJA addition. For the time-course experiment, we analysed the expression of 10 terpene synthase homologues found in the grape EST database by real time PCR. The expression of at least 4 of these putative sesquiterpenes synthase genes are induced by MeJA, with maximum expression at 24 hours, therefore 48 hours before the peak of sesquiterpene accumulation.
Flavour Biosynthesis Pathways in Grape Cell Cultures: Sesquiterpenes Biosynthesis
D'ONOFRIO, CLAUDIO;
2009-01-01
Abstract
The aim of this study was to discover genes involved in grape flavour pathways by elicitation of grape cell suspension. Among numerous elicitors tested, methyl jasmonate (MeJA) and jasmonic acid (JA) induce a wide range of sesquiterpenes in ‘Cabernet Sauvignon’ and Riesling berry cell suspension cultures. Sesquiterpenes, the largest subclass of natural terpenes compounds, may be synthesised by two different pathways: the mevalonate pathways (MVA) localised in the cytosol and the methylerytritol phoshate pathway (DOXP/MEP) localised in the plastids. Using specific inhibitors we found that in our cell suspension MeJA induced sequiterpenes are synthesised by the MVA pathway. A set of experiments performed to optimize sesquiterpenes production, indicated that the efficacy of MeJA concentration is strictly dependent upon the packed cell volume (PCV) of cell suspension at the moment of elicitor addition. In time course studies it was found that the peak in sesquiterpene accumulation was recorded 72 hours after MeJA addition. For the time-course experiment, we analysed the expression of 10 terpene synthase homologues found in the grape EST database by real time PCR. The expression of at least 4 of these putative sesquiterpenes synthase genes are induced by MeJA, with maximum expression at 24 hours, therefore 48 hours before the peak of sesquiterpene accumulation.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
