Effects of organochlorines on microbial diversity and community structure in Phragmites australis rhizosphere.

This study investigated the impacts of an organochlorine (OC, γ-hexachlorocyclohexane and chlorobenzenes) mixture on microbial communities associated to Phragmites australis rhizosphere. Seventy-eight distinct colony morphotypes were isolated, cultivated and analysed by 16S rDNA sequence analysis. Toxicity tests confirmed sensitivity (e.g. Hevizibacter, Acidovorax) or tolerance (e.g. Bacillus, Aeromonas, Pseudomonas, Sphingomonas) of isolates. Rhizosphere analysis by pyrosequencing showed the microbial adaptation induced by OC exposure. Among the most abundant molecular operational taxonomic units, 80 % appeared to be tolerant (55 % opportunist, 25 % unaffected) and 20 % sensitive. P. australis rhizosphere exposed to OCs was dominated by phylotypes related to α-, ß- and γ-Proteobacteria. Specific genera were identified which were previously described as chlorinated organic pollutant degraders: Sphingomonas sp., Pseudomonas sp., Devosia sp. and Sphingobium sp. P. australis could be suitable plants to maintain their rhizosphere active microbial population which can tolerate OCs and potentially improve the OC remediation process in part by biodegradation.

Biological responses of maize (Zea mays) plants exposed to chlorobenzenes. Case study of monochloro-, 1,4-dichloro- and 1,2,4-trichloro-benzenes.

A 7-day-exposure time experiment was designed to investigate the phytotoxicity of chlorobenzenes (CBs) on Zea mays seedlings, focusing on the growth and generation of oxidative stress. Significant growth inhibition (based on biomass gain) was observed for exposure to monochlorobenzene (MCB), dichlorobenzene (DCB) and trichlorobenzene (TCB) concentrations higher than 10 mg l(-1). It would seem that CBs inhibit cell division, since the mitotic index decreased for roots exposed to DCB at 80 mg l(-1) dose (8%) and to all the TCB concentrations tested (20% inhibition). CBs exposure resulting in an increase in the oxidative stress response in maize seedlings [reactive oxygen species like H(2)O(2), antioxidant enzymes (POD, GR), lipid peroxidation] correlated to the compound's degree of chlorination, where damage increasing with the number of chlorine atoms (MCB < DCB < TCB). This biological response was also dependent on the dose-exposure. Z. mays exposed to CBs at concentrations <10 mg l(-1) did not induce sufficient oxidative damage to cause root cell death. Therefore, CBs at current environmental concentrations are unlikely to produce evident phytotoxic effects on Z. mays seedlings.

Cell cycle disruption and apoptosis as mechanisms of toxicity of organochlorines in Zea mays roots.

Organochlorine pesticides (OCPs) are widespread environmental pollutants; two of them are highly persistent: lindane (γHCH) and chlordecone (CLD). Maize plants cope with high levels of OCP-environmental pollution, however little is known about cellular mechanisms involved in plant response to such OCP-exposures. This research was aimed at understanding the physiological pathways involved in the plant response to OCPs in function of a gradient of exposure. Here we provide the evidences that OCPs might disrupt root cell cycle leading to a rise in the level of polyploidy possibly through mechanisms of endoreduplication. In addition, low-to-high doses of γHCH were able to induce an accumulation of H2O2 without modifying NO contents, while CLD modulated neither H2O2 nor NO production. [Ca(2+)]cytosolic, the caspase-3-like activity as well as TUNEL-positive nuclei and IP-positive cells increased after exposure to low-to-high doses of OCPs. These data strongly suggest a cascade mechanism of the OCP-induced toxic effect, notably with an increase in [Ca(2+)]cytosolic and caspase-3-like activity, suggesting the activation of programmed cell death pathway.

A comparative study on the uptake and translocation of organochlorines by Phragmites australis.

Organochlorines (OCs) are persistent chemicals found in various environmental compartments. The differences in the uptake of (14)C-labeled 1,4-dichlorobenzene (DCB), 1,2,4-trichlorobenzene (TCB) and γ-hexachlorocyclohexane (γHCH) by Phragmites australis were investigated under hydroponic conditions. The first step in sorption appears to be correlated with the hydrophobic nature of the compounds, since log-linear correlations were obtained between root concentration factor and partition coefficient (LogK(ow)). After 7 days of exposure, plant uptake of DCB, TCB, γHCH was significant with bioconcentration factors reaching 14, 19 and 15, respectively. Afterwards, uptake and translocation were seen to be more complex, with a loss of the simple relationship between uptake and LogK(ow). Linear correlations between the bioconcentration/translocation factors and the physico-chemical properties of OCs were shown, demonstrating that translocation from roots to shoots increases with solubility and volatility of the OCs. This suggests that OC-translocation inside plants might result from the combination of two processes, xylem sap flow and vapor fluxes. (14)C-phytovolatilization was measured and was correlated with the volatility of the compounds; the more volatile OCs being most the likely to be phytovolatilized from foliar surfaces (p=0.0008). Thus, OC-uptake/translocation appears to proceed at a rate that depends mostly on the OCs hydrophobicity, solubility and volatility.

Response of phase II detoxification enzymes in Phragmites australis plants exposed to organochlorines.

Mixed pollution is a characteristic of many industrial sites and constructed wetlands. Plants possessing an enzymatic detoxifying system that is able to handle xenobiotics seems to be a viable option for the removal of mixed persistent contaminants such organochlorines (OCs: monochlorobenzene (MCB), 1,4-dichlorobenzene (DCB), 1,2,4-trichlorobenzene (TCB), γ-hexachlorocyclohexane (HCH)). In this study, Phragmites australis plants were exposed to sub-lethal concentrations of OCs (7 days), in single-exposure (0.8 to 10 mg l(-1)) and in mixture of OCs (0.2 mg l(-1) MCB + 0.2 mg l(-1) DCB + 2.5 mg l(-1) TCB + 0.175 mg l(-1) HCH). Studies were conducted on the detoxification phase II enzymes; glutathione S-transferases (GST), and glucosyltransferases (UGT). Measurements of GST and UGT activities revealed that OCs may be buffered by glutathione and glucose conjugation. There appeared to be a correlation between the effects on phase II enzymes and the degree of chlorination of the benzene ring with, for example, the greatest effects being obtained for HCH exposure. In the case of mixed pollution, the induction of some GST isoenzymes (CDNB, 35 % non-significant) and UGT (118 %) in leaves and the inhibition of phase II enzymes in the other organs were measured. UGTs appear to be key enzymes in the detoxification of OCs.

Concentration responses to organochlorines in Phragmites australis.

Phragmites australis shows potential for the phytoremediation of chlorinated chemicals. Also there has been some attempt to determine the phytotoxic effects of organochlorines (OC). This study reports for lindane (HCH), monochlorobenzene (MCB), 1,4-dichlorobenzene (DCB) and 1,2,4-trichlorobenzene (TCB), a no-observed-effect-concentration (NOEC(7d)) that was 1000-300,000 times higher than environmental concentrations. Nevertheless, the combined OC mixture (NOEC(7d) level of each congener) induced a synergistic toxic effect, causing a severe drop (70%) in chlorophyll concentration. The mixture 0.2 mg L(-1) MCB+0.2 mg L(-1) DCB+2.5 mg L(-1) TCB+0.175 mg L(-1) HCH, that was 15 times more concentrated than environmental OC mixture, did not cause phytotoxicity during 21 days. Antioxidant enzymes were affected immediately after the start of exposure (3 days), but the plants showed no signs of stress thereafter. These data suggest that environmental OC mixtures do not pose a significant risk to P. australis.