Molecular cloning and expression of a gene encoding alcohol acyltransferase (MdAAT2) from apple (cv. Golden Delicious).

Volatile esters are major aroma components of apple, and an alcohol acyltransferase (AAT) catalyzes the final step in ester biosynthesis. The gene MdAAT2, which encodes a predicted 51.2 kDa protein containing features of other acyl transferases, was isolated from Malus domestica Borkh. (cv. Golden Delicious). In contrast to other apple varieties, the MdAAT2 gene of Golden Delicious is exclusively expressed in the fruit. The MdAAT2 protein is about 47.9 kDa and mainly localized in the fruit peel, as indicated by immunoblot and immunolocalization analysis. Northern blot and immunoblot analysis showed that the transcription and translation of MdAAT2 have a positive correlation with apple AAT enzyme activity and ester production, except in the later ripening stage, suggesting that MdAAT2 is involved in the regulation of ester biosysthesis and that a post-translation modification may be involved in regulation of AAT enzyme activity. Tissue disk assays of fruit peel revealed that using extraneous alcohols can recover the corresponding ester formation. Transcription and translation of MdAAT2 were both depressed by 1-methylcyclopropene (1-MCP) treatment and subsequent ester production was also prevented. These results suggest that: (1) ester production is mainly regulated by MdAAT2; (2) ethylene is also involved in this regulatory progress and (3) ester compounds rely principally on the availability of substrates.