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Determination of ornithine, putrescine, N‐methylputrescine and N‐methylpyrroline pools in tobacco tissue by high‐performance liquid chromatography
Author(s) -
Feth Friedhelm,
Wagner Karl G.
Publication year - 1989
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1989.tb02065.x
Subject(s) - putrescine , chromatography , metabolite , ornithine , chemistry , derivatization , high performance liquid chromatography , dansyl chloride , ornithine decarboxylase , enzyme , biochemistry , arginine , amino acid
A procedure for the determination of metabolites of the biochemical pathway ornithine to N‐methyl‐δ 1 ‐pyrrolinium salt (N‐methylpyrroline) is described. Plant tissue was extracted with 0.5 M HCl and the extract purified on C 18 ‐cartridges. Ornithine was reacted with o ‐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.