Determination of ornithine, putrescine, N‐methylputrescine and N‐methylpyrroline pools in tobacco tissue by high‐performance liquid chromatography

Publisher Website
Google Scholar
Abstract
A procedure for the determination of metabolites of the biochemical pathway ornithine to N‐methyl‐δ1‐pyrrolinium salt (N‐methylpyrroline) is described. Plant tissue was extracted with 0.5MHCl and the extract purified on C18‐cartridges. Ornithine was reacted witho‐phthaldialdehyde, putrescine and N‐methylputrescine with dansyl chloride and the products were separated by reversed‐phase high‐performance liquid chromatography (HPLC). N‐methylpyrroline was determined by cation‐exchange HPLC without derivatization. The metabolites in the roots of tobacco (Nicotiana) species with different nicotine‐producing capacities were determined. Furthermore, the specific activities of the enzymes ornithine decarboxylase (EC 4.1.1.17), putrescine N‐methyltransferase (EC 2.1.1.53) and N‐methylputrescine oxidase were determined. Both the metabolite pools and the enzyme activities were correlated with the different nicotine‐producing capacities of the different tobacco species.