- Sulfur containing aroma volatiles are important contributors to the distinctive aroma of melon and other fruits. Melon cultivars and accessions differ in the content of sulfur-containing and other volatiles. L-Methionine has been postulated to serve as a precursor of these volatiles. Incubation of melon fruit cubes with (13) C- and (2) H- labeled L-methionine revealed two distinct catabolic routes into volatiles: one apparently involving the action of an L-methionine aminotransferase and preserving the main carbon skeleton of L-methionine. The second route apparently involves the action of an L-methionine-γ-lyase activity, releasing methanethiol, a backbone for the formation of thiol-derived aroma volatiles. Exogenous L-methionine also generated non-sulfur volatiles by further metabolism of α-ketobutyrate, a product of L-methionine-γ-lyase activity. α-Ketobutyrate was further metabolized into L-isoleucine and to other important melon volatiles including non-sulfur branched and straight-chain esters. Cell-free extracts derived from ripe melon fruit exhibited L-methionine-γ-lyase enzymatic activity. A melon gene (CmMGL), ectopically expressed in E. coli was shown to encode a protein possessing L-methionine-γ-lyase enzymatic activity. The expression of CmMGL was relatively low in early stages of melon fruit development, but increased in the flesh of ripe fruits depending on the cultivar tested. Moreover, the levels of expression of CmMGL in recombinant inbred lines co-segregated with the levels of the S-volatiles enriched with +1 m/z unit and postulated to be produced via this route. Our results indicate that L-methionine is a precursor of both sulfur and non-sulfur aroma volatiles in melon fruit. © 2013 The Authors. The Plant Journal © 2013 Blackwell Publishing Ltd.