Control of Cortaderia selloana with a glyphosate-based herbicide led to a short-term stimulation of soil fungal communities.

In the north of Spain, Cortaderia selloana plants have invaded ecosystems of high ecological value. Control of this species is carried out with the application of glyphosate-based formulations. The aim of this work was to determine, under microcosm conditions, the short-term (2 months) effects of the application of a glyphosate-based herbicide (Roundup®) on C. selloana rhizosphere microbial communities. To this purpose, before and after the application of Roundup®, several parameters that provide information on the biomass, activity and diversity of rhizosphere fungal and bacterial communities (enzyme activities, basal and substrate-induced respiration, potentially mineralizable nitrogen, nitrification potential rate, ergosterol content and community-level profiles with Biolog™ plates and ARISA) were determined. We observed a stimulation of some microbial parameters, in particular those related to fungal communities. Further research is needed to determine the long-term consequences of this short-term fungal stimulation for soil functioning.

Evaluation of the phytostabilisation efficiency in a trace elements contaminated soil using soil health indicators.

The efficiency of a remediation strategy was evaluated in a mine soil highly contaminated with trace elements (TEs) by microbiological, ecotoxicological and physicochemical parameters of the soil and soil solution (extracted in situ), as a novel and integrative methodology for assessing recovery of soil health. A 2.5-year field phytostabilisation experiment was carried out using olive mill-waste compost, pig slurry and hydrated lime as amendments, and a native halophytic shrub (Atriplex halimus L.). Comparing with non-treated soil, the addition of the amendments increased soil pH and reduced TEs availability, favoured the development of a sustainable vegetation cover (especially the organic materials), stimulated soil microorganisms (increasing microbial biomass, activity and functional diversity, and reducing stress) and reduced direct and indirect soil toxicity (i.e., its potential associated risks). Therefore, under semi-arid conditions, the use of compost and pig slurry with A. halimus is an effective phytostabilisation strategy to improve soil health of nutrient-poor soils with high TEs concentrations, by improving the habitat function of the soil ecosystem, the reactivation of the biogeochemical cycles of essential nutrients, and the reduction of TEs dissemination and their environmental impact.

Plant tolerance to diesel minimizes its impact on soil microbial characteristics during rhizoremediation of diesel-contaminated soils.

Soil contamination due to petroleum-derived products is an important environmental problem. We assessed the impacts of diesel oil on plants (Trifolium repens and Lolium perenne) and soil microbial community characteristics within the context of the rhizoremediation of contaminated soils. For this purpose, a diesel fuel spill on a grassland soil was simulated under pot conditions at a dose of 12,000 mg diesel kg(-1) DW soil. Thirty days after diesel addition, T. repens (white clover) and L. perenne (perennial ryegrass) were sown in the pots and grown under greenhouse conditions (temperature 25/18 °C day/night, relative humidity 60/80% day/night and a photosynthetic photon flux density of 400 µmol photon m(-2) s(-1)) for 5 months. A parallel set of unplanted pots was also included. Concentrations of n-alkanes in soil were determined as an indicator of diesel degradation. Seedling germination, plant growth, maximal photochemical efficiency of photosystem II (F(v)/F(m)), pigment composition and lipophylic antioxidant content were determined to assess the impacts of diesel on the studied plants. Soil microbial community characteristics, such as enzyme and community-level physiological profiles, were also determined and used to calculate the soil quality index (SQI). The presence of plants had a stimulatory effect on soil microbial activity. L. perenne was far more tolerant to diesel contamination than T. repens. Diesel contamination affected soil microbial characteristics, although its impact was less pronounced in the rhizosphere of L. perenne. Rhizoremediation with T. repens and L. perenne resulted in a similar reduction of total n-alkanes concentration. However, values of the soil microbial parameters and the SQI showed that the more tolerant species (L. perenne) was able to better maintain its rhizosphere characteristics when growing in diesel-contaminated soil, suggesting a better soil health. We concluded that plant tolerance is of crucial importance for the recovery of soil health during rhizoremediation of contaminated soils.

Native plant communities in an abandoned Pb-Zn mining area of northern Spain: implications for phytoremediation and germplasm preservation.

Plants growing on metalliferous soils from abandoned mines are unique because of their ability to cope with high metal levels in soil. In this study, we characterized plants and soils from an abandoned Pb-Zn mine in the Basque Country (northern Spain). Soil in this area proved to be deficient in major macronutrients and to contain toxic levels of Cd, Pb, and Zn. Spontaneously growing native plants (belonging to 31 species, 28 genera, and 15 families) were botanically identified. Plant shoots and rhizosphere soil were sampled at several sites in the mine, and analyzed for Pb, Zn and Cd concentration. Zinc showed the highest concentrations in shoots, followed by Pb and Cd. Highest Zn concentrations in shoots were found in the Zn-Cd hyperaccumulator Thlaspi caerulescens (mean = 18,254 mg Zn kg(-1) DW). Different metal tolerance and accumulation patterns were observed among the studied plant species, thus offering a wide germplasm assortment for the suitable selection of phytoremediation technologies. This study highlights the importance of preserving metalliferous environments as they shelter a unique and highly valuable metallicolous biodiversity.

Phytostabilization of metal contaminated soils.

The contamination of soils with heavy metals represents a worldwide environmental problem of great concern. Traditional methods for the remediation of metal contaminated soils are usually very expensive and frequently induce adverse effects on soil properties and biological activity. Consequently, biological methods of soil remediation like phytoremediation (the use of green plants to clean up contaminated sites) are currently receiving a great deal of attention. In particular, chemophytostabilization of metal contaminated soils (the use of metal tolerant plants together with different amendments like organic materials, liming agents, or phosphorus compounds and such) to reduce metal mobility and bioavailability in soils appears most promising for sites contaminated with high levels of several metals when phytoextraction is not a feasible option. During chemophytostabilization processes, one must at all times be cautious with a possible future reversal of soil metal immobilization, with concomitant adverse environmental consequences.

Phytoextraction potential of two Rumex acetosa L. accessions collected from metalliferous and non-metalliferous sites: effect of fertilization.

Metal tolerance and phytoextraction potential of two common sorrel (Rumex acetosa L.) accessions, collected from a Pb/Zn contaminated site (CS, Lanestosa) and an uncontaminated site (UCS, Larrauri), were studied in fertilized and non-fertilized pots prepared by combining soil samples from both sites in different proportions (i.e., 0%, 33%, 66% and 100% of Lanestosa contaminated soil). The original metalliferous mine soil contained 20480, 4950 and 14 mg kg(-1) of Zn, Pb and Cd, respectively. The microcosm experiment was carried out for two months under greenhouse controlled conditions. It was found that fertilization increased mean plant biomass of both accessions as well as their tolerance. However, only the CS accession survived all treatments even though its biomass decreased proportionally according to the percentage of contaminated mine soil present in the pots. This metallicolous accession would be useful for the revegetation and phytostabilization of mine soils. Due to its high concentration and bioavailability in the contaminated soil, the highest values of metal phytoextracted corresponded to Zn. The CS accession was capable of efficiently phytoextracting metal from the 100% mine soil, indeed reaching very promising phytoextraction rates in the fertilized pots (6.8 mg plant(-1) month(-1)), similar to the ones obtained with hyperaccumulator plants. It was concluded that fertilization is certainly worth being considered for phytoextraction and revegetation with native plants from metalliferous soils.

Dendroremediation of heavy metal polluted soils.

Heavy metals are among the most common and harmful pollutants reaching the soil ecosystem all over the world. Phytoextraction is an effective, non-intrusive, inexpensive, aesthetically pleasing, socially accepted, highly promising phytotechnology for the remediation of soils polluted with heavy metals. To overcome the so-called 'Achilles' heel' of phytoextraction, namely, the long time needed for effective remediation, this phytotechnology should be combined with other profit-making activities such as forestry or bioenergy production. Dendroremediation, or the use of trees to clean up polluted soil and water, appears of great potential for metal phytoextraction, especially when using fast-growing tree species, for example, willows (Salix sp. pl.) and poplars (Populus sp. pl.). Most important, the ecologic and environmental risks of dispersing heavy metals into the ecosystems by dendroremediation strategies should be minimized by selecting the right tree species, properly managing/disposing the polluted plant material, or a combination of both options.

Synthesis of low molecular weight thiols in response to Cd exposure in Thlaspi caerulescens.

In this study, we investigated the accumulation of phytochelatins (PCs) and other low molecular weight (LMW) thiols in response to Cd exposure in two contrasting ecotypes differing in Cd accumulation. Using a root elongation test, we found that the highly accumulating ecotype Ganges was more tolerant to Cd than the low Cd-accumulation ecotype Prayon. L-buthionine-(S,R)-sulphoximine (BSO), a potent inhibitor of the gamma-glutamylcysteine synthetase gamma-ECS) (an enzyme involved in the PC biosynthetic pathway), increased the Cd sensitivity of Prayon, but had no effect on Ganges. Although PC accumulation increased in response to Cd exposure, no significant differences were observed between the two ecotypes. Cd exposure induced a dose-dependent accumulation of both Cys and a still unidentified LMW thiol in roots of both ecotypes. Root accumulation of Cys and this thiol was higher in Ganges than in Prayon; the ecotypic differences were more pronounced when the plants were treated with BSO. These findings suggest that PCs do not contribute to the Cd hypertolerance displayed by the Ganges ecotype of Thlaspi caerulescens, whereas Cys and other LMW thiols might be involved.

Bioluminescent bacterial biosensors for the assessment of metal toxicity and bioavailability in soils.

A major factor governing the toxicity of heavy metals in soils is their bioavailability. Traditionally, sequential extraction procedures using different extractants followed by chemical analysis have been used for determining the biologically available fraction of metals in soils. Yet, the transfer of results obtained on non-biological systems to biological ones is certainly questionable. Therefore, bioluminescence-based bacterial biosensors have been developed using genetically engineered microorganisms, constructed by fusing transcriptionally active components of metal resistance mechanisms to lux genes from naturally bioluminescent bacteria like Vibrio fischeri for the assessment of metal toxicity and bioavailability in polluted soils. As compared to chemical methods, bacterial biosensors present certain advantages, such as selectivity, sensitivity, simplicity, and low cost. Despite certain inherent limitations, bacterial bioluminescent systems have proven their usefulness in soils under laboratory and field conditions. Finally, green fluorescent protein-based bacterial biosensors are also applicable for determining with high sensitivity the bioavailability of heavy metals in soil samples.

Phytoextraction and phytofiltration of arsenic.

Arsenic, a ubiquitous contaminant in groundwater and soils, is currently drawing much public attention. Arsenic-contaminated soils can be cleaned up via phytoextraction-the use of plants to extract the arsenic from soil and transport it into aboveground tissues. Arsenic removal from polluted soils can be carried out using hyperaccumulator ferns like the Chinese brake fern Pteris vittata, which accumulates very high concentrations of the element in aboveground tissues. The capacity of the plant to take up large concentrations of arsenic, even at low levels in soil, illustrates efficient bioaccumulation. The possibility of using Pteris ferns to remove arsenic from water by phytofiltration has been proposed.

Climbing a ladder: a step-by-step approach to understanding the concept of agroecosystem health.

Population and individual health is linked to agroecosystem health. To comprehend the concept of agroecosystem health, one should climb a ladder consisting of several successive steps, each rung presenting a certain degree of instability (conceptual difficulty and uncertainty) in an advisable but not inevitable order. Here we suggest a ladder consisting of the following concepts: ecosystem, agroecosystem, biodiversity, sustainability, ecosystem health, and agroecosystem health. Although these concepts are to a certain extent well understood and grasped by scientists, politicians, natural resource managers, and environmentalists, some steps are still highly debatable, unclear, and present a considerable degree of reluctance to be defined and understood. Consequently, much empirical and theoretical effort must be made to construct solid conceptual ladders made up of such steps. In this enterprise, a traditional reductionistic approach confining interpretations to narrow scientific disciplines is unadvisable. Holistic, transdisciplinary approaches are required to reach the desired goal.

Phytoremediation of organic contaminants in soils.

Soil pollution, a very important environmental problem, has been attracting considerable public attention over the last decades. Unfortunately, the enormous costs associated with the removal of pollutants from soils by means of traditional physicochemical methods have been encouraging companies to ignore the problem. Phytoremediation is an emerging technology that uses plants to clean up pollutants in the environment. As overwhelmingly positive results have become available regarding the ability of plants to degrade certain organic compounds, more and more people are getting involved in the phytoremediation of organic contaminants. Phytoremediation of organics appears a very promising technology for the removal of these contaminants from polluted sites.

Phytoextraction: a cost-effective plant-based technology for the removal of metals from the environment.

Phytoremediation is an emerging technology that uses plants to clean up pollutants (metals and organics) from the environment. Within this field of phytoremediation, the utilization of plants to transport and concentrate metals from the soil into the harvestable parts of roots and above-ground shoots is usually called phytoextraction. Most traditional remediation methods do not provide acceptable solutions for the removal of metals from soils. By contrast, phytoextraction of metals is a cost-effective approach that uses metal-accumulating plants to clean up these soils. Subsequently, the harvestable parts, rich in accumulated metals, can be easily and safely processed by drying, ashing or composting. Some extracted metals can also be reclaimed from the ash, generating recycling revenues. Phytoextraction appears a very promising technology for the removal of metal pollutants from the environment and may be, at present, approaching commercialization.

Rhodobacter capsulatus DNA topoisomerase I purification and characterization.

A 30-kDa DNA topoisomerase has been purified to near homogeneity from the purple nonsulfur photosynthetic bacterium Rhodobacter capsulatus. The enzyme is recognized by an antibody against a 16-mer peptide sequence from human DNA topoisomerase I. The purified enzyme is a type I topoisomerase. Consistent with the properties of other prokaryotic type I DNA topoisomerases, the isolated enzyme is unable to relax positively supercoiled DNA and absolutely requires divalent cations for its relaxation activity. However, regardless of the Mg+2 concentrations, ATP concentrations above 5 mM completely inhibit the relaxing activity. The enzyme is sensitive to high salt concentrations and the optimal activity occurs at salt concentrations between 3 and 30 mM for monovalent cations. Single-stranded M13 DNA is a strong inhibitor of this relaxing activity. The enzyme is inhibited by ethidium bromide, confirming that this DNA topoisomerase is incapable of relaxing positive supercoils. Topoisomerase I-specific inhibitors like Hoechst 32258 and actinomycin D inhibit the enzymatic activity while the enzyme is resistant to type II topoisomerase inhibitors such as norfloxacin, nalidixic acid, and novobiocin. From these enzymatic characteristics, we conclude that the R. capsulatus DNA topoisomerase is a prokaryotic type I DNA topoisomerase.

Morphological and biochemical responses of Bacillus subtilis to selenite stress.

When introduced into a chemically defined minimal medium supplemented with 1 mM sodium selenite (79 ppm Se(o)), Bacillus subtilis was found to undergo a series of morphological and biochemical adaptations. The morphological changes included the formation of "round bodies" associated with the detoxification of selenite to elemental selenium. Round bodies observed transiently were not apparent during balanced growth of cells adapted previously to selenite-containing medium. Under balanced growth conditions, cell structures similar to "round bodies", could be produced by treating cells with lysozyme. The selenite-induced structural alterations in cells were accompanied by an increase in the content of thioredoxin and the associated enzyme, NADP-thioredoxin reductase. The results suggest that the biovalence transformation of high levels of selenite may involve a dithiol system.

Aerobic chromate reduction by Bacillus subtilis.

We have studied the reduction of hexavalent chromium (chromate) to the less toxic trivalent form by using cell suspensions and cell-free extracts from the common soil bacterium, Bacillus subtilis. B. subtilis was able to grow and reduce chromate at concentrations ranging from 0.1 to 1 mM K2CrO4. Chromate reduction was not affected by a 20-fold excess of nitrate-compound that serves as alternate electron acceptor and antagonizes chromate reduction by anaerobic bacteria. Metabolic poisons including sodium azide and sodium cyanide inhibited chromate reduction. Reduction was effected by a constitutive system associated with the soluble protein fraction and not with the membrane fraction. The reducing activity was heat labile and showed a Km of 188 microns CrO4(2)-. The reductase can mediate the transfer of electrons from NAD(P)H to chromate. The results suggest that chromate is reduced via a detoxification system rather than dissimilatory electron transport.

Selenite bioremediation potential of indigenous microorganisms from industrial activated sludge.

Ten bacterial strains were isolated from the activated sludge waste treatment system (BIOX) at the Exxon refinery in Benicia, California. Half of these isolates could be grown in minimal medium. When tested for selenite detoxification capability, these five isolates (members of the genera Bacillus, Pseudomonas, Enterobacter and Aeromonas), were capable of detoxifying selenite with kinetics similar to those of a well characterized Bacillus subtilis strain (168 Trp+) studied previously. The selenite detoxification phenotype of the Exxon isolates was stable to repeated transfer on culture media which did not contain selenium. Microorganisms isolated from the Exxon BIOX reactor were capable of detoxifying selenite. Treatability studies using the whole BIOX microbial community were also carried out to evaluate substrates for their ability to support growth and selenite bioremediation. Under the appropriate conditions, indigenous microbial communities are capable of remediating selenite in situ.

Bioavailability of selenium accumulated by selenite-reducing bacteria.

The bioavailability of selenium (Se) was determined in bacterial strains that reduce selenite to red elemental Se (SeO). A laboratory strain of Bacillus subtilis and a bacterial rod isolated from soil in the vicinity of the Kesterson Reservoir, San Joaquin Valley, CA, (Microbacterium arborescens) were cultured in the presence of 1 mM sodium selenite (Na2SeO3). After harvest, the washed, lyophilized B. Subtilis and M. arborescens samples contained 2.62 and 4.23% total Se, respectively, which was shown to consist, within error, entirely of SeO. These preparations were fed to chicks as supplements to a low-Se, vitamin E-free diet. Three experiments showed that the Se in both bacteria had bioavailabilities of approx 2% that of selenite. A fourth experiment revealed that gray SeO had a bioavailability of 2% of selenite, but that the bioavailability of red SeO depended on the way it was prepared (by reduction of selenite). When glutathione was the reductant, bioavailability resembled that of gray SeO and bacterial Se; when ascorbate was the reductant, bioavailability was twice that level (3-4%). These findings suggest that aerobic bacteria such as B. subtilis and M. arborescens may be useful for the bioremediation of Se-contaminated sites, i.e., by converting selenite to a form of Se with very low bioavailability.

Physiological mechanisms regulating the conversion of selenite to elemental selenium by Bacillus subtilis.

We have demonstrated that the common soil bacterium, Bacillus subtilis, reduces selenite to an insoluble and much less toxic product--the red form of elemental selenium. Reduction was effected by an inducible system that appears to deposit elemental selenium between the cell wall and the plasma membrane. Glucose and sucrose supported selenite reduction. Although malate and citrate supported growth, no significant reduction of selenite occurred, indicating the importance of the redox state of the culture substrate. Selenite reduction in the millimolar concentration range (i.e., cultures supplemented with 1 mM selenite) was not affected by a ten-fold excess of nitrate or sulfate--compounds that serve as alternate electron acceptors and antagonize selenite reduction by anaerobic bacteria. Similarly, nitrite and sulfite did not significantly affect the rate or extent of selenite reduction. B.subtilis was able to grow and produce selenium (Se degree) at selenite concentrations ranging from 0.6 microM to 5 mM (50 ppb to 395 ppm selenium). At the lowest selenite concentration tested, 50 ppb selenium, B.subtilis removed 95% of the selenite from the liquid phase. The results suggest that selenite is reduced via an inducible detoxification system rather than dissimilatory electron transport. The findings establish the potential utility of B.subtilis for the bioremediation of selenite-polluted sites.