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The role of 1-pyrroline was studied via feeding experiments using rice calli cultures to gain further insight into the key steps of 2-acetyl-1-pyrroline (2AP) biosynthesis in rice. The origin of the acetyl donor was also studied through stable isotope labelled substrates. Incubation of fresh calli from a fragrant rice variety (Aychade) and a non-fragrant variety (Gladio×Fidji K2) with 1-pyrroline led to a significant increase in 2AP in both varieties. Importantly, the amount of 2AP in the non-fragrant variety could be greatly enhanced by this supplementation. When rice calli were fed with increasing levels of 1-pyrroline, 2AP levels increased accordingly. Our data show that 1-pyrroline is a limiting factor for 2AP synthesis in rice. Heat treatment of calli suggested that 1-pyrroline might be enzymatically acetylated. The presence of labelled 2AP in calli supplemented with [U-(13)C]glucose, sodium acetate (1,2-(13)C2) and sodium octanoate (1,2,3,4-(13)C4) suggested that these compounds are possible candidates for acetyl group-donors of 2AP, predominately in the form of intact labelled (13)C2-units., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

In greenhouse experiments, Aychade, a fragrant rice variety, was grown under one level of salt solution (EC of 3800 ± 400 μS·cm(-1)) sufficient to induce salt stress in rice. Timing and duration of salt solution application varied according to the growth stages. 2-Acetyl-1-pyrroline (2AP), a characteristic flavor compound of fragrant rice as well as biogenetically related compounds, proline, and γ-aminobutyric acid (GABA) were quantified. Salt treatments induced 2AP synthesis in the leaves, but the increase was often higher in the vegetative phase. This increase was correlated with proline level but not with that of GABA. Interestingly the grains from all the salt treated plants contained significantly higher levels of 2AP (733-998 μg·kg(-1)) than those from the control (592 μg·kg(-1)). The highest 2AP synthesis occurred when the plants were subjected to salt treatment during whole vegetative or reproductive phases. However in the latter case crop yield decreased significantly.