Growth parameters influencing uptake of chlordecone by Miscanthus species.

Because of its high persistence in soils, t1/2=30years, chlordecone (CLD) was classified as a persistent organic pollutant (POP) by the Stockholm Convention in 2009.The distribution of CLD over time has been heterogeneous, ranging from banana plantations to watersheds, and contaminating all environmental compartments. The aims of this study were to (i) evaluate the potential of Miscanthus species to extract chlordecone from contaminated soils, (ii) identify the growth parameters that influence the transfer of CLD from the soil to aboveground plant parts. CLD uptake was investigated in two species of Miscanthus, C4 plants adapted to tropical climates. M. sinensis and M.×giganteus were transplanted in a soil spiked with [14C]CLD at environmental concentrations (1mgkg-1) under controlled conditions. Root-shoot transfer of CLD was compared in the two species after two growing periods (2 then 6months) after transplantation. CLD was found in all plant organs, roots, rhizomes, stems, leaves, and even flower spikes. The highest concentration of CLD was in the roots, 5398±1636 (M.×giganteus) and 14842±3210ngg-1 DW (M. sinensis), whereas the concentration in shoots was lower, 152±28 (M.×giganteus) and 266±70ngg-1 DW (M. sinensis) in soil contaminated at 1mgkg-1. CLD translocation led to an acropetal gradient from the bottom to the top of the plants. CLD concentrations were also monitored over two complete growing periods (10months) in M. sinensis grown in 8.05mgkg-1 CLD contaminated soils. Concentrations decreased in M. sinensis shoots after the second growth period due to the increase in organic matters in the vicinity of the roots. Results showed that, owing to their respective biomass production, the two species were equally efficient at phytoextraction of CLD.

Chlordecone Transfer and Distribution in Maize Shoots.

Chlordecone (CLD) is a persistent organic pollutant (POP) that was mainly used as an insecticide against banana weevils in the French West Indies (1972-1993). Transfer of CLD via the food chain is now the major mechanism for exposure of the population to CLD. The uptake and the transfer of CLD were investigated in shoots of maize, a C4 model plant growing under tropical climates, to estimate the exposure of livestock via feed. Maize plants were grown on soils contaminated with [(14)C]CLD under controlled conditions. The greatest part of the radioactivity was associated with roots, nearly 95%, but CLD was detected in whole shoots, concentrations in old leaves being higher than those in young ones. CLD was thus transferred from the base toward the plant top, forming an acropetal gradient of contaminant. In contrast, results evidenced the existence of a basipetal gradient of CLD concentration within leaves whose extremities accumulated larger amounts of CLD because of evapotranspiration localization. Extractable residues accounted for two-thirds of total residues both in roots and in shoots. This study highlighted the fact that the distribution of CLD contamination within grasses resulted from a conjunction between the age and evapotranspiration rate of tissues. CLD accumulation in fodder may be the main route of exposure for livestock.

Uptake and distribution of chlordecone in radish: different contamination routes in edible roots.

Chlordecone (CLD) was an organochlorine insecticide mainly used to struggle against banana weevils in the French West Indies. Forbidden since 1993, it has been a long-term contaminant of soils and aquatic environments. Crops growing in contaminated soils lead to human exposure by food consumption. We used radiolabeled [(14)C]-CLD to investigate the contamination ways into radish, a model of edible roots. Radish plants were able to accumulate CLD in both roots (RCF35d 647) and tubers (edible parts, CF35d 6.3). CLD was also translocated to leaves (CF35d 1.7). The contamination of tuber was mainly due to peridermic adsorption or CLD systemic translocation to the pith. TSCF was 3.44×10(-)(3). CLD diffused across periderm to internal tissues. We calculated a mean flux of diffusion J through periderm about 5.71×10(-)(14)gcm(-)(2)s(-)(1). We highlighted different contamination routes of the tuber, (i) adsorption on periderm followed by diffusion of CLD towards underlying tissues, cortex, xylem, and pith (ii) adsorption by roots and translocation by the transpiration stream followed by diffusion from xylem vessels towards inner tissues, pith, and peripheral tissues, cortex and periderm. Concerning chemical risk assessment for other tubers, contamination would depend on various parameters, the thickness of periderm and CLD periderm permeance, the origin of secondary tissues - from cortex and/or pith - , the importance of xylem flow in tuber, and the lipid amount within tuber.

Metabolic fate of 2,4-dichlorophenol and related plant residues in rats.

This study compared the metabolic fate of [(14)C]-DCP, [(14)C]-residues from radish plants, and purified [(14)C]-DCP-(acetyl)glucose following oral administration in rats. A rapid excretion of radioactivity in urine occurred for [(14)C]-DCP, [(14)C]-DCP-(acetyl)glucose, and soluble residues, 69, 85, and 69% within 48 h, respectively. Radio-HPLC profiles of 0-24 h urine from rats fed [(14)C]-DCP and [(14)C]-DCP-(acetyl)glucose were close and qualitatively similar to those obtained from plant residues. No trace of native plant residues was detected under the study conditions. The structures of the two major peaks were identified by MS as the glucuronide and the sulfate conjugates of DCP. The characterization of a dehydrated glucuronide conjugate by MS and NMR of DCP was unusual. In contrast to soluble residues, bound residues were mainly excreted in feces, 90% within 48 h, whereas total residues were eliminated in both urine and feces. For total residues, the radioactivity in feces was higher than expected from the percentage of soluble and bound residues in radish plants. This result highlighted that less absorption took place when residues were present in the plant matrix as compared to plant-free residues and DCP.

Metabolic fate of [¹4C]diuron and [¹4C]linuron in wheat (Triticum aestivum) and radish (Raphanus sativus).

Metabolism of xenobiotics in plants usually occurs in three phases, phase I (primary metabolism), phase II (conjugation processes), and phase III (storage). The uptake and metabolism of [(14)C]diuron and [(14)C]linuron were investigated in wheat and radish. Seeds were sown in quartz sand and irrigated with a nutrient solution of either radioactive herbicide. Plants were harvested after two weeks, and metabolites were extracted and then analyzed by radio-reverse-high-performance liquid chromatography (HPLC). Uptake of the two molecules was higher in radish compared to wheat. Translocation of parent compounds and related metabolites from roots to aerial plant parts was important, especially for radish. A large proportion of extractable residues were found in radish whereas nonextractable residues amounted to 30% in wheat, mainly associated with roots. Chemical structure of metabolites was thereafter identified by acid, alkaline, and enzymatic hydrolyses followed by electrospray ionization mass spectrometry (ESI-MS) and proton nuclear magnetic resonance spectroscopy ((1)H NMR). This study highlighted the presence of diuron and linuron metabolites conjugated to sugars in addition to N-demethylation and N-demethoxylation products.

Metabolic fate of [14C] chlorophenols in radish (Raphanus sativus), lettuce (Lactuca sativa), and spinach (Spinacia oleracea).

Chlorophenols are potentially harmful pollutants that are found in numerous natural and agricultural systems. Plants are a sink for xenobiotics, which occur either intentionally or not, as they are unable to eliminate them although they generally metabolize them into less toxic compounds. The metabolic fate of [ (14)C] 4-chlorophenol (4-CP), [ (14)C] 2,4-dichlorophenol (2,4-DCP), and [ (14)C] 2,4,5-trichlorophenol (2,4,5-TCP) was investigated in lettuce, spinach, and radish to locate putative toxic metabolites that could become bioavailable to food chains. Radish plants were grown on sand for four weeks before roots were dipped in a solution of radiolabeled chlorophenol. The leaves of six-week old lettuce and spinach were treated. Three weeks after treatments, metabolites from edible plant parts were extracted and analyzed by high performance liquid chromatography (HPLC) and characterized by mass spectrometry (MS), and nuclear magnetic resonance spectroscopy (NMR). Characterization of compounds highlighted the presence of complex glycosides. Upon hydrolysis in the digestive tract of animals or humans, these conjugates could return to the toxic parent compound, and this should be kept in mind for registration studies.

Metabolic fate of [(14)C]-2,4-dichlorophenol in tobacco cell suspension cultures.

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.

Metabolism of [14C]-2,4-dichlorophenol in edible plants.

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.

Uptake and metabolic fate of [14C]-2,4-dichlorophenol and [14C]-2,4-dichloroaniline in wheat (Triticum aestivum) and soybean (Glycine max).

The uptake and metabolism of [14C]-2,4-dichlorophenol (DCP) and [14C]-2,4-dichloroaniline (DCA) were investigated in wheat and soybean. Seeds were exposed to a nutrient solution containing 50 microM of one of two radiolabeled compounds, and plant organs were harvested separately after 18 days of growth. In wheat, uptake of [14C]-2,4-DCP was 16.67 +/- 2.65 and 15.50 +/- 2.60% of [14C]-2,4-DCA. In soybean, uptake of [14C]-2,4-DCP was significantly higher than [14C]-2,4-DCA uptake, 38.39 +/- 2.56 and 18.98 +/- 1.64%, respectively. In the case of [14C]-2,4-DCP, the radioactivity absorbed by both species was found mainly associated with roots, whereas [14C]-2,4-DCA and related metabolites were associated with aerial parts, especially in soybean. In wheat, nonextractable residues represented 7.8 and 8.7% of the applied radioactivity in the case of [14C]-2,4-DCP and [14C]-2,4-DCA, respectively. In soybean, nonextractable residues amounted to 11.8 and 5.8% of the total radioactivity for [14C]-2,4-DCP and [14C]-2,4-DCA, respectively. In wheat, nonextractable residues were nearly equivalent to extractable residues for [14C]-2,4-DCP, whereas they were greater for [14C]-2,4-DCA. In soybean, the amount of extractable residues was significantly greater for both chemicals. However, in both species, nonextractable residues were mainly associated with roots. Isolation of soluble residues was next undertaken using excised shoots (wheat) or excised fully expanded leaves including petioles (soybean). Identification of metabolite structures was made by comparison with authentic standards, by enzymatic hydrolyses, and by electrospray ionization-mass spectrometric analyses. Both plant species shared a common metabolism for [14C]-2,4-DCP and [14C]-2,4-DCA since the malonylated glucoside conjugates were found as the final major metabolites.