Application Of Separated Leaf Cell Suspension To Xenobiotic Metabolism In Plant

Metabolic profiles of C-14-labeled primary metabolites from several pesticides, 4-cyanophenol (1), 3-phenoxybenzoic acid (2), 3-phenoxybenzyl alcohol (3), 3,5-dichloroaniline (4), and (1RS)-trans-2,2dimethyl-3-(2-methylprop-1-enyl)cyclopropanecarboxylic acid (5), were examined by using enzymatically separated leaf cell suspension from seedlings of cabbage (Brassica oleracea) and tomato (Lycopersicon esculentum). After 1 day of incubation, the metabolites were extensively transformed in cabbage, whereas they were scarcely metabolized in tomato. The major metabolic pathways were the phase II reactions leading to a number of conjugates such as glucoside/malonylglucoside of 1-5, malate of 2, and glutamate of 4. The oxidation of 1 and 2 was observed as a minor reaction to produce 4-hydroxybezoic acid and 3-(4-hydroxyphenoxy)benzoic acid. The chemical identities of the secondary metabolites were determined by various spectrometric analyses (LC-MS, LC-MS/MS, and NMR) and/or HPLC cochromatography with the synthetic reference standards. As a result, this separated leaf cell suspension system was found to well reproduce the in vivo plant metabolism.