The metabolism of 2,4,6-trinitrotoluene (TNT) was investigated in tobacco cell suspension cultures amended with [14C]-TNT. Five metabolites were purified and characterized. Temporal evolution of metabolites was monitored during a 120 h incubation period. Metabolites structure was identified by acid and enzymatic hydrolysis, followed by electrospray ionization mass spectrometry and 1H and 13C NMR spectroscopy analyses. The majority of metabolites were conjugates formed by glycose conjugation on the hydroxylamine group of either 2-hydroxylamino-4,6-dinitrotoluene (2-HADNT) or 4-hydroxylamino-2,6-dinitrotoluene (4-HADNT), which led to monoglycoside then to diglycoside. Various diglycosides were observed with gentiobioside or sophoroside formation. Bound residues represented a small fraction (<10% of initial 14C) irrespective of the interval after TNT amendment. Free ADNT was detected only in the medium. This study highlights the central role played by HADNT in the TNT metabolic pathway in tobacco cell suspension culture, and the key role of these compounds and of glycosyltransferases in TNT phytoremediation processes.

The fate of imidacloprid was investigated in tomato plants during 75 days in soil contaminated by 14 C-imidacloprid. Leaves and fruits were separately analysed for total radioactivity and metabolites. Almost 85% of plant radioactivity was translocated to shoots. Radioactivity concentrations decreased from bottom leaves to top leaves. Desnitro-imidacloprid was the main metabolite in leaves. Nevertheless, more than 50% of the leave radioactivity corresponded to imidacloprid. Residue concentrations were similar in all fruits (62.9 ng g -1 ), irrespective of their position on plant. In fruits more than 85% of the radioactivity was due to imidacloprid. The small fraction of residues translocated to fruits depended on the low xylem flow in fruits.

In plant tissues, xenobiotics often are conjugated with natural constituents such as sugars, amino acids, glutathione, and malonic acid. Usually, conjugation processes result in a decrease in the reactivity and toxicity of xenobiotics by increasing the water solubility and polarity of conjugates, and reducing their mobility. Due to their lack of an efficient excretory system, xenobiotic conjugates finally are sequestered in plant storage compartments or cell vacuoles, or are integrated as bound residues in cell walls. Chlorophenols are potentially harmful pollutants that are found in numerous natural and agricultural systems. We studied the metabolic fate of 2,4-dichlorophenol (DCP) in cell-suspension cultures of tobacco (Nicotiana tabacum L.). After a standard metabolism experiment, 48 h of incubation with a [U-phenyl-(14)C]-DCP solution, aqueous extracts of cell suspension cultures were analyzed by high-performance liquid chromatography (HPLC). Metabolites then were isolated and their chemical structures determined by enzymatic and chemical hydrolyses, electrospray ionization-mass spectrometry in negative mode (ESI-NI), and (1)H nuclear magnetic resonance analyses. The main terminal metabolites identified were DCP-glycoside conjugates, DCP-(6-O-malonyl)-glucoside, DCP-(6-O-acetyl)-glucoside, and their precursor, DCP-glucoside. More unusual and complex DCP conjugates such as an alpha(1-->6)-glucosyl-pentose and a triglycoside containing a glucuronic acid were further characterized. All the metabolites identified were complex glycoside conjugates. However, these conjugates still may be a source of DCP in hydrolysis reactions caused by microorganisms in the environment or in the digestive tract of animals and humans. Removal of xenobiotics by glycoside conjugation thus may result in underestimation of the risk associated with toxic compounds like DCP in the environment or in the food chain.

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Several 2,4-dichlorophenoxyacetic acid (2,4-D)-sensitive plants have been modified by genetic engineering with tfdA gene to acquire 2,4-D tolerance. The expression product of this gene degrades 2,4-D to 2,4-dichlorophenol (DCP), which is less phytotoxic but could cause a problem of food safety. After a comparison of 2,4-D and DCP metabolism in transgenic 2,4-D-tolerant and wild cotton (Gossypium hirsutum L.), a direct study of DCP metabolism in edible plants was performed. After petiolar uptake of a [U-phenyl-(14)C]-DCP solution followed by a 48 h water chase, aqueous extracts were analysed by high-performance liquid chromatography. Metabolites were thereafter isolated and their structural identities were determined by enzymatic and chemical hydrolyses and mass spectrometry analyses. The metabolic fate of DCP was equivalent to 2,4-D metabolism in transgenic 2,4-D-tolerant cotton. In addition, DCP metabolism was similar in transgenic and wild cotton. The major terminal metabolites were DCP-saccharide conjugates in all species, essentially DCP-(6-O-malonyl)-glucoside or its precursor DCP-glucose. The significance of this metabolic pathway with regard to food safety is discussed.