Partitioning of New Carbon as 11C in Nicotiana tabacum Reveals Insight into Methyl Jasmonate Induced Changes in Metabolism

We examined the timeline by which methyl jasmonate (MeJA) reprograms new carbon partitioning into key metabolite pools. The radioactive isotope C-11 (t(1/2) 20.4 min), administered to intact leaves of Nicotiana tabacum L. (cv Samsun) as (CO2)-C-11 gas enabled us to measure changes in new carbon partitioning into soluble sugar and amino acid pools of [C-11]photosynthate. A 500 mu M MeJA treatment resulted in a decrease in the [C-11]soluble sugar pool and an increase in the [C-11]amino acid pool after 4 h. This pattern was more pronounced 15 h after treatment. We also examined the timeline for C-11-partitioning into aromatic amino acid metabolites of the shikimate pathway. [C-11]Tyrosine, [C-11]phenylalanine and [C-11]tryptophan were elevated 1.5-fold, 12-fold and 12-fold, respectively, relative to controls, 4 h after MeJA treatment, while endogeneous pools were unchanged. This suggests that only new carbon is utilized during early stages of defense induction. By 15 h, [C-11]tyrosine and [C-11]phenylalanine returned to baseline while [C-11]tryptophan was elevated 30-fold, suggesting that MeJA exerts selective control over the shikimate pathway. Finally, we measured trans-cinnamic acid levels as a gauge of downstream phenolic metabolism. Levels were unchanged 4 h after MeJA treatment relative to controls, but were increased 2-fold by 15 h, indicating a lag in response of secondary metabolism.