Radiotracer computer modeling evidence that phospho-base methylation is the main route of choline synthesis in tobacco
Among flowering plants, the synthesis of choline (Cho) from ethanolamine (EA) can potentially occur via three parallel, interconnected pathways involving methylation of free bases, phospho-bases, or phosphatidyl-bases. The authors investigated which pathways operate in tobacco (Nicotiana tabacum L.) because previous work has shown that the endogenous Cho supply limits accumulation of glycine betaine in transgenic tobacco plants engineered to convert Cho to glycine betaine. The kinetics of metabolite labeling were monitored in leaf discs supplied with [{sup 33}P]phospho-EA,[{sup 33}P]phospho-monomethylethanolamine, or [{sup 14}C]formate, and the data were subjected to computer modeling. Because partial hydrolysis of phospho-bases occurred in the apoplast, modeling of phospho-base metabolism required consideration of the re-entry of [{sup 33}P]phosphate into the network. Modeling of [{sup 14}C]formate metabolism required consideration of the labeling of the EA and methyl moieties of Cho. Results supported the following conclusions: (a) The first methylation step occurs solely at the phospho-base level; (b) the second and third methylations occur mainly (83%--92% and 65%--85%, respectively) at the phospho-base level, with the remainder occurring at the phosphatidyl-base level; and (c) free Cho originates predominantly from phosphatidylcholine rather than from phospho-Cho. This study illustrates how computer modeling of radiotracer data, in conjunction with information on chemical pool sizes, can provide a coherent, quantitative picture of fluxes within a complex metabolic network.
- Research Organization:
- Univ. of Florida, Gainesville, FL (US)
- OSTI ID:
- 20062532
- Journal Information:
- Plant Physiology (Bethesda), Vol. 123, Issue 1; Other Information: PBD: May 2000; ISSN 0032-0889
- Country of Publication:
- United States
- Language:
- English
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