How Plants Cope with Foreign Compounds. Translocation of xenobiotic glutathione conjugates in roots of barley (Hordeum vulgare) (9 pp)

Background, Aim and Scope Numerous herbicides and xenobiotic organic pollutants are detoxified in plants to glutathione conjugates. Following this enzyme catalyzed reaction, xenobiotic GS-conjugates are thought to be compartmentalized in the vacuole of plant cells. In the present study, evidence is presented for long range transport of these conjugates in plants, rather than storage in the vacuole. To our knowledge this is the first report about the unidirectional long range transport of xenobiotic conjugates in plants and the exudation of a glutathione conjugate from the root tips. This could mean that plants possess an excretion system for unwanted compounds which give them similar advantages as animals. Materials and Methods: Barley plants (Hordeum vulgare L. cv. Cherie) were grown in Petri dishes soaked with tap water in the greenhouse. - Fluorescence Microscopy. Monobromo- and Monochlorobimane, two model xenobiotics that are conjugated rapidly in plant cells with glutathione, hereby forming fluorescent metabolites, were used as markers for our experiments. Their transport in the root could be followed sensitively with very good temporal and spatial resolution. Roots of barley seedlings were cut under water and the end at which xenobiotics were applied was fixed in an aperture with a thin latex foil and transferred into a drop of water on a cover slide. The cover slide was fixed in a measuring chamber on the stage of an inverse fluorescence microscope (Zeiss Axiovert 100). - Spectrometric enzyme assay. Glutathione S-transferase (GST) activity was determined in the protein extracts following established methods. Aliquots of the enzyme extract were incubated with 1-chloro-2,4-dinitrobenzene (CDNB), or monochlorobimane. Controls lacking enzyme or GSH were measured. - Pitman chamber experiments. Ten days old barley plants or detached roots were inserted into special incubation chambers, either complete with tips or decapitated, as well as 10 days old barley plants without root tips. Compartment A was filled with a transport medium and GSH conjugate or L-cysteine conjugate. Compartments B and C contained sugar free media. Samples were taken from the root tip containing compartment C and the amount of conjugate transported was determined spectro-photometrically. Results: The transport in roots is unidirectional towards the root tips and leads to exsudation of the conjugates at rates between 20 and 200 nmol min-1. The microscopic studies have been complemented by transport studies in small root chambers and spectroscopic quantification of dinitrobenzene-conjugates. The latter experiments confirm the microscopic studies. Furthermore it was shown that glutathione conjugates are transported at higher rates than cysteine conjugates, despite of their higher molecular weights. This observation points to the existence of glutathione specific carriers and a specific role of glutathione in the root. Discussion: It can be assumed that long distance transport of glutathione conjugates within the plant proceeds like GSH or amino acid transport in both, phloem and xylem. The high velocity of this translocation of the GS-X is indicative of an active transport. For free glutathione, a rapid transport-system is essential because an accumulation of GSH in the root tip inhibits further uptake of sulfur. Taking into account that all described MRP transporters and also the GSH plasmalemma ATPases have side activities for glutathione derivatives and conjugates, co-transport of these xenobiotic metabolites seems credible. - On the other hand, when GS-B was applied to the root tips from the outside, no significant uptake was observed. Thus it can be concluded that only those conjugates can be transported in the xylem which are formed inside the root apex. Having left the root once, there seems to be no return into the root vessels, probably because of a lack of inward directed transporters. Conclusions: Plants seem to possess the capability to store glutathione conjugates in the vacuole, but under certain conditions, these metabolites might also undergo long range transport, predominantly into the plant root. The transport seems dependent on specific carriers and is unidirectional, this means that xenobiotic conjugates from the rhizosphere are not taken up again. The exudation of xenobiotic metabolites offers an opportunity to avoid the accumulation of such compounds in the plant. Recommendations and Perspectives: The role of glutathione and glutathione related metabolites in the rhizosphere has not been studied in any detail, and only scattered data are available on interactions between the plant root and rhizosphere bacteria that encounter such conjugates. The final fate of these compounds in the root zone has also not been addressed so far. It will be interesting to study effects of the exuded metabolites on the biology of rhizosphere bacteria and fungi.     

Closing a Loop: Substance Flow Analysis of Nitrogen and Phosphorus in the Rainbow Trout Production and Domestic Consumption System in Finland

Ongoing eutrophication is changing the Baltic Sea ecosystem. Aquaculture causes relatively small-scale nutrient emissions, but local environmental impact may be considerable. We used substance flow analysis (SFA) to identify and quantify the most significant flows and stocks of nitrogen (N) and phosphorus (P) related to rainbow trout aquaculture in Finland. In 2004–2007, the input of nutrients to the system in the form of fish feed was 829 t N year⁻¹ and 115 t P year⁻¹. Around one-fifth of these nutrients ended up as food for human consumption. Of the primary input, 70% ended up in the Baltic Sea, directly from aquaculture and indirectly through waste management. The nutrient cycle could be closed partially by using local fish instead of imported fish in rainbow trout feed, thus reducing the net load of N and P to a fraction.     

Investigation of the accumulation of ash,heavy metals,and polycyclic aromatic hydrocarbons to assess the stability of lysis–cryptic growth sludge reduction in sequencing batch reactor

The accumulation of ash, heavy metals, and polycyclic aromatic hydrocarbons (collectively called potential accumulating substances, PAS) was evaluated to ascertain the stability of lysis–cryptic growth sludge reduction process (LSRP) for municipal sludge treatment. One sequencing batch reactor (SBR) incorporated with homogenization was run to test the LSRP and another SBR as a control. The continuous monitoring results for 2 months showed that the ash and heavy metals slightly increased, and the polycyclic aromatic hydrocarbons decreased by 18.0%, indicating that there may be negligible accumulations during the LSRP. Their accumulations met pattern I, as demonstrated by statistical analysis, proving no PAS accumulation for LSRP. This was further confirmed by sludge activity and system performance. Moreover, the mechanism for no PAS accumulation was discussed. It was concluded that the LSRP was stable with no worries about PAS accumulation under the operational conditions.     

DNA damage in oral epithelial cells of individuals chronically exposed to indoor radon (<Superscript>222</Superscript>Rn) in a hydrothermal area

Hydrothermal areas are potentially hazardous to humans as volcanic gases such as radon (²²²Rn) are continuously released from soil diffuse degassing. Exposure to radon is estimated to be the second leading cause of lung cancer, but little is known about radon health-associated risks in hydrothermal regions. This cross-sectional study was designed to evaluate the DNA damage in the buccal epithelial cells of individuals chronically exposed to indoor radon in a volcanic area (Furnas volcano, Azores, Portugal) with a hydrothermal system. Buccal epithelial cells were collected from 33 individuals inhabiting the hydrothermal area (Ribeira Quente village) and from 49 individuals inhabiting a non-hydrothermal area (Ponta Delgada city). Indoor radon was measured with Ramon 2.2 detectors. Chromosome damage was measured by micronucleus cytome assay, and RAPD-PCR was used as a complementary tool to evaluate DNA damage, using three 10-mer primers (D11, F1 and F12). Indoor radon concentration correlated positively with the frequency of micronucleated cells (r _s = 0.325, p = 0.003). Exposure to radon is a risk factor for the occurrence micronucleated cells in the inhabitants of the hydrothermal area (RR = 1.71; 95% CI, 1.2–2.4; p = 0.003). One RAPD-PCR primer (F12) produced differences in the banding pattern, a fact that can indicate its potential for detecting radon-induced specific genomic alterations. The observed association between chronic exposure to indoor radon and the occurrence of chromosome damage in human oral epithelial cells evidences the usefulness of biological surveillance to assess mutations involved in pre-carcinogenesis in hydrothermal areas, reinforcing the need for further studies with human populations living in these areas.     

Cosmogenic exposure dating constraints for coastal landslide evolution on the Island of Malta (Mediterranean Sea)

Landslides affecting the north-western coast of the Island of Malta have been investigated and monitored for 10 years. As a result of a bathymetric survey, it was discovered the deposits continued out onto the seafloor, thus raising questions as to the timing of their development. Furthermore it was uncertain as to which environment they developed in and which factors controlled their movements. The aim of this paper is to investigate representative detachments to chronologically constrain these mass movement events and outline their spatial and temporal evolution. Samples for exposure dating using the cosmogenic nuclide ³⁶Cl were collected from head scarps and blocks located within two long-term monitored landslides characterised by extensive block slides. The results indicate the oldest dated block detachment occurring in a subaerial environment at ca. 21 ka, when the sea level was about 130 m lower than at present. Mass movement possibly accelerated when sea level reached the landslide toe during the post-glacial marine transgression. Considering the timing of block movement, the landslide deposits observed today appear to be related to a first-time failure involving a large part of the slope, though and alternative hypothesis is also taken here into account. This main event is likely to have been followed by secondary movements influenced by toe undercutting and clay saturation due to rising sea level. However, further research on mass movement kinematics is required in order to model their evolution and explore whether this interpretation is widely applicable along the Maltese coast.     

Simultaneous adsorption of Remazol brilliant blue and Disperse orange dyes on red mud and isotherms for the mixed dye system

The paper presents the adsorption of Remazol brilliant blue (RBB) and Disperse orange 25 (DO25) dyes from aqueous solution of the mixture of dyes onto concentrated sulphuric acid-treated red mud (ATRM). First-order derivative spectrophotometric method was developed for the analysis of RBB and DO25 in mixed dye aqueous solution to overcome the limitations arising due to interference in the zero-order spectral method. The optimum conditions to maximize RBB adsorption favoured the adsorption of RBB, and those for DO25 favoured DO25 adsorption from the mixed dye aqueous solutions. Presence of a second dye always inhibited the adsorption of a target dye. The uptake and percentage adsorption of each of the dyes onto ATRM from the aqueous solution of the mixture of dyes decreased considerably with increasing concentrations of the other dye showing the antagonistic effect. Monocomponent Langmuir isotherm fitted the mixed dye adsorption equilibrium data better than the monocomponent Freundlich isotherm. However, monocomponent models are suitable for the fixed concentration of the other dye. Modified Langmuir isotherm model adequately predicted the multi-component adsorption equilibrium data for RBB-DO25-ATRM adsorption system with a good accuracy and is more generic from the application point of view.     

Pesticide removal from waste spray-tank water by organoclay adsorption after field application: an approach for a formulation of cyprodinil containing antifoaming/defoaming agents

Purpose  

Many reports on purification of water containing pesticides are based on studies using unformulated active ingredients. However, most commercial formulations contain additives/adjuvants or are manufactured using microencapsulation which may influence the purification process. Therefore, the main objective of this work was to develop and test a pilot scheme for decontaminating water containing pesticides formulated with antifoaming/defoaming agents.