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.     

Photocatalytic and combined anaerobic-photocatalytic treatment of textile dyes

A photocatalytic process based on immobilized titanium dioxide was used to treat crude solutions of azo, anthraquinone and phthalocyanine textile dyes. In addition, the process was applied to the treat autoxidized chemically reduced azo dyes, i.e. representatives of recalcitrant dye residues after biological sequential anaerobic-aerobic treatment. Photocatalysis was able to remove more than 90% color from crude as well as autoxidized chemically reduced dye solutions. UV-absorbance and COD were also removed but to a lower extent (50% in average). The end products of photocatalytic treatment were not toxic toward methanogenic bacteria. The results demonstrate that photocatalysis can be used as a pre- or post-treatment method to biological anaerobic treatment of dye-containing textile wastewater.     

Lipid metabolism of the Antarctic krill Euphausia superba and its ecological implications

 Various developmental stages (early larvae to adults) of Euphausia superba have been collected in different seasons in the Weddell Sea, the Lazarev Sea and off the Antarctic Peninsula to investigate the role of lipids and fatty acids in the life cycle of the Antarctic krill. The total-lipid data for E. superba exhibited seasonal variations, with low lipid levels in late winter/early spring and the highest levels in autumn. Seasonal changes were most pronounced in the immature and adult specimens, increasing from about 10% lipid of dry mass to more than 40%. The fatty-acid compositions of the younger stages were dominated by 20:5(n-3), 22:6(n-3) and 16:0. These are typical phospholipid fatty acids, which are major biomembrane constituents. The phospholipid composition was similar in the older stages. With increasing storage of triacylglycerols in the lipid-rich immature and adult stages, the fatty acids 14:0, 16:0 and 18:1(n-9) prevailed, comprising about 70% of total triacylglycerol fatty acids. The trophic-marker fatty acids 16:1(n-7) and 18:4(n-3), indicating phytoplankton ingestion, were less abundant. They reflected, however, the dependence of the larvae on phytoplankton as well as the seasonal changes in algal composition. The generally close linear relationships between fatty acids and lipid suggest that the fatty-acid compositions of the collected specimens were largely independent of the respective developmental stage, season and region. The linear fit indicates that triacylglycerol accumulation started at a level of about 5% of total lipid. Considering the various overwintering scenarios under discussion, the life cycle and reproductive strategies of krill are discussed in the context of the lipid metabolism and fatty-acid composition of E. superba. Lipid production is effective enough to accumulate large energy reserves for the dark season, but E. superba does not exhibit the sophisticated biosynthetic pathways known from other Antarctic euphausiids and copepods. Although important, lipid utilisation appears to be just one of several strategies of E. superba to thrive under the extreme Antarctic conditions, and this pronounced versatility may explain the success of this species in the Southern Ocean. Received: 16 June 2000 / Accepted: 18 December 2000     

Differences in herbivore preferences, phlorotannin production, and nutritional quality between juvenile and adult tissues from marine brown algae

 Juvenile and adult marine organisms differ in their morphology, chemistry, physiology, behavior, and ecology. Because juvenile algae are thinner, smaller, and have more delicate tissues than adults, they are often assumed to be more susceptible to grazers. We examined within-species food preferences of four common generalist herbivores for juvenile and adult tissues of eight common brown algae in two-choice laboratory food-preference experiments. Our results showed that juvenile algae did not tend to be a preferred food of herbivores. Juvenile tissues were significantly preferred over adult tissues in only four of the 32 combinations of algae and herbivores tested. In 12 experiments, adult tissues were preferred over juvenile tissues, and no choice occurred in the remaining 16 experiments. When sea urchins exhibited a preference, it was always for adult tissues. The other three herbivores, an isopod and two snails, were more variable in their choices, sometimes preferring juveniles, sometimes adults, and sometimes having no preference. We measured nitrogen and phlorotannin concentrations in adult and juvenile seaweeds to see whether these parameters were correlated with herbivore food preferences. Nitrogen levels were similar in juveniles and adults of three algal species and were higher in juveniles of two. Phlorotannin levels were higher in juveniles of four species and lower in juveniles of one. The other three species showed no differences in phlorotannin levels. Phlorotannin concentrations decreased with increasing juvenile size in three species and increased with increasing size in one species. Neither nitrogen nor phlorotannin concentrations explained overall herbivore food preferences for algae of different stages. Our results suggest that preferences of certain grazers for juvenile algae are not as strong as previously assumed and are dependent on herbivore species. Preferences between juveniles and adults are likely to be determined by a combination of morphological and chemical features of the tissues and the unique responses of herbivore species to those features. Received: 10 April 2000 / Accepted: 19 November 2000     

Dimethylsulfoniopropionate and dimethylsulfide in the marine flatworm Convoluta roscoffensis and its algal symbiont

 A distinct smell of dimethylsulfide (DMS) was noted at the edge of the intertidal mudflat of Marennes-Oléron Bay, at the French Atlantic coast, where dense populations of the marine flatworm Convoluta roscoffensis Graff (Platyhelminthes: Turbellaria) were present. DMS is the cleavage product of dimethylsulfoniopropionate (DMSP). DMSP was shown to be present in high amounts in sediment containing the flatworm as well as in axenic cultures of the symbiotic alga Tetraselmis sp. that was isolated from the flatworm. In untreated sediment samples containing C. roscoffensis the concentration of DMS was as high as ∼55 μmol l⁻¹ sediment, and in samples that were fixed with glutaraldehyde the concentration of DMS was even three orders of magnitude higher (∼66 mmol l⁻¹ sediment). This rapid cleavage of DMSP to DMS in fixed samples was unexpected. Pure DMSP was stable in glutaraldehyde, and it was therefore concluded that a DMSP-lyase was responsible for cleavage in the field samples. The isolated symbiotic alga, Tetraselmis sp., did not show DMSP-lyase activity, indicating that DMSP-lyase may have been present in the flatworm, although the role of bacteria could not be excluded. The Chl a-specific DMSP content of C. roscoffensis (∼200 mmol g⁻¹) was much higher than that of Tetraselmis sp. (∼30 mmol g⁻¹). Possibly, DMSP was not only present in the symbiotic alga, but was also incorporated in the body tissue of the flatworm. It remains unclear what the function of DMSP is in C. roscoffensis. In Tetraselmis sp., but not in C. roscoffensis, DMSP increased with increasing salinity. It was concluded that salinity probably does not play an important role in the dynamics of DMSP and DMS in sediment containing C. roscoffensis. Received: 21 January 2000 / Accepted: 29 August 2000     

Metal concentrations in American oyster <Emphasis Type="Italic">Crassotrea virginica</Emphasis> and adjacent sediments from harvestable and non-harvestable sites in the Southeastern USA

Human population growth in coastal areas continues to threaten estuarine ecosystems and resources. Populations of Crassostrea virginica have declined across the USA due to water quality degradation, disease pressure, alteration of habitat, and other changes related to anthropogenic impacts. Metals that may be present in estuarine habitats can bioaccumulate in oysters, with potential consequences to the health of oysters and humans consumers. This study (1) evaluated the occurrence and relationships of metal concentrations in oyster tissue versus estuarine sediments, (2) examined oyster tissue concentrations in relation to state water quality designations, and (3) evaluated the potential risk for humans from oyster consumption related to metal concentrations from harvestable waters. Results indicated metal concentrations in sediments and oysters along coastal South Carolina remain low compared to other areas and that concentrations in oyster tissue and adjacent sediments were not highly correlated with each other. However, high concentrations of some metals occurred in oysters sampled from areas designated as Approved for Harvesting. This is important because most harvest area designation systems rely on regular bacterial monitoring when evaluating the safety of consumption. Others safety measurements may be necessary as part of routine monitoring.     

Human factors and tidal influences on water quality of an urban river in Can Tho, a major city of the Mekong Delta, Vietnam

In this study, we focused on water quality in an urban canal and the Mekong River in the city of Can Tho, a central municipality of the Mekong Delta region, southern Vietnam. Water temperature, pH, electrical conductivity, BOD₅, COD_Cr, Na⁺, Cl^?, NH₄ ⁺?N, SO₄ ^2??S, NO₃ ^??N, and NO₂ ^??N for both canal and river, and tide level of the urban canal, were monitored once per month from May 2010 to April 2012. The urban canal is subject to severe anthropogenic contamination, owing to poor sewage treatment. In general, water quality in the canal exhibited strong tidal variation, poorer at lower tides and better at higher tides. Some anomalies were observed, with degraded water quality under some high-tide conditions. These were associated with flow from the upstream residential area. Therefore, it was concluded that water quality in the urban canal changed with a balance between dilution effects and extent of contaminant supply, both driven by tidal fluctuations in the Mekong River.     

Concentrations of organophosphate esters and brominated flame retardants in German indoor dust samples

While it is known that the ingestion of indoor dust contributes substantially to human exposure to the recently restricted polybrominated diphenyl ethers (PBDEs), the situation for one class of potential replacements, i.e. organophosphate esters (OPEs), used in a variety of applications including as flame retardants has yet to be fully characterised. In this study, surface dust from twelve different cars from various locations throughout Germany were analysed for eight OPEs, decabromodiphenyl ethane (DBDPE), and eight PBDEs. In five cars, tris-(1,3-dichloro-2-propyl) phosphate (TDCPP) was the dominant compound with concentrations up to 620 μg g(-1) dust. High concentrations of tri-cresyl phosphate (TCP) (up to 150 μg g(-1)) were also detected in two samples of car dust. Dust from ten offices in the same building in Ludwigsburg, Germany was also analysed. In these samples, tri (2-butoxyethyl) phosphate (TBEP) predominated with an average concentration of 7.0 μg g(-1) dust, followed by tris (1-chloro-2-propyl) phosphate (TCPP) at 3.0 μg g(-1) and triphenyl phosphate (TPhP) at 2.5 μg g(-1) dust. Although caution must be exercised given the relatively small database reported here; this study provides evidence that cars and offices from Germany are significantly more contaminated with OPEs than PBDEs. Average concentrations of ΣOPEs were ten times higher in car than in office dust. This is the first study to provide data on a wide range of OPE concentrations in German indoor dust samples.