<i>In Vitro</i> Metabolism of Fluoroimide in the Cell Suspension of Apple Leaves

Abstract

In vitro metabolism of fluoroimide was studied in isolated apple cell suspension culture. The cells were isolated from freshly excised apple leaves by Macerozyme R-10, and cultured in Murashige-Skoog medium containing 1mg/l of 2, 4-D at 27°C in the dark. When 14C-fluoroimide (labeled at ethylene carbons) in DMSO solution was added to the cell suspension medium, fluoroimide was readily hydrolyzed and incorporated into the cells. The major metabolites were 4-fluoro-dichloromaleanilic acid (12), 2, 3-dichloromaleic acid (15) and (E)-2, 3-dichloroacrylic acid (14), and these were found both in the cell extracts and the medium. These In vitro metabolic profiles were similar to those of the intact Inu-apple tree, but in this study was identified (E)-2, 3-dichloroacrylic acid (14) by its isolation from massive cell culture of non-labeled fluoroimide. The growth inhibition activity of the apple cells was observed in fluoroimide and two kinds of dechlorinated metabolites (2 and 5) which were slightly produced.

Application of Separated Leaf Cell Suspension to Xenobiotic Metabolism in Plant

“…The formation and decline of metabolites were examined in detail for [ 14 C]-5 using the cabbage leaf cell suspension with exposure periods of 1, 2, and 4 days. The metabolic distribution reached a plateau within 1 day, and approximately 100% of the radioactivity detected in the cabbage leaf cell was the malonylglucose conjugate of 5 (15). On the basis of these data, the incubation period in the study examining metabolic profiles of [ 14 C]-1-5 was determined to be 1 day after treatment.…”

“…Interestingly, metabolites of 1 and 2 in the leaf cell suspension were detected in the culture medium, which facilitated their isolation and purification for spectroscopic analyses. A number of researchers have reported that significant amounts of polar metabolites are excreted to the extracellular media from the suspension culture cells − and separated leaf cells . Recently, the membrane transport of the plant secondary metabolites has been extensively studied and the transporters and/or channels of plants are shown as deeply involved in the membrane transport of alkaloids, terpenoids, phenol, and wax in plant cell.…”

“…The following gradient systems were used for the typical analysis, separation, and purification of each metabolite: 0. 14), and 23.5 (15).…”

“…For example, it has been shown that these cells are capable of performing processes such as photosynthesis, respiration, and syntheses of RNA, protein, and lipid − . However, sufficient investigation has not been conducted to examine the metabolic features of xenobiotics using this method . From this standpoint, we have conducted metabolism studies using the separated leaf cell suspension of cabbage (Brassica oleracea) and tomato (Lycopersicon esculentum) with phenolic, alcoholic, anilinic, and carboxylic compounds, which are the primary metabolites from various kinds of agrochemicals.…”

Metabolism of the persistent plasticizer chemical bis(2-ethylhexyl) phthalate in cell suspension cultures of wheat (Triticum aestivum L.). Discrepancy from the intact plant

“…It is widely assumed that cultured plant cells can serve as a metabolic model of the intact plant. In fact, a considerable similarity between metabolism in cell cultures and in the intact plant has been observed for example for 2,4-D (Mumma and Hamilton, 1979;Bristol et al, 1977; Scheel and Sandermann, 1981), diphenamide (Davis et al, 1978), cisanilide (Frear and Swanson, 1975), fluorodifen (Locke and Baron, 1972), carbaryl (Locke et al, 1976), diclofopmethyl (Dusky et al, 1980(Dusky et al, , 1982, delmonem and Mumma, 1982), fenvalerate (Davidonis and Mumma, 1984), fluoroimide (Ohori and Aizawa, 1983), baythroid (Preiss et al, 1984), 4-chloro-2-methylphenoxyacetic acid (Cole and Loughman, 1983), and pentachlorophenol (Scheel et al, 1984).…”