María Balseiro-Romero

Enhanced Degradation of Diesel in the Rhizosphere of after Inoculation with Diesel-Degrading and Plant Growth-Promoting Bacterial Strains.

The association of plants and rhizospheric bacteria provides a successful strategy to clean up contaminated soils. The purpose of this work was to enhance diesel degradation in rhizosphere by inoculation with selected bacterial strains: a diesel degrader (D), plant growth-promoting (PGP) strains, or a combination (D+PGP). Plants were set up in pots with the A or B horizon of an umbric Cambisol (A and B) spiked with diesel (1.25%, w/w). After 1 mo, the dissipation of diesel range organics (DRO) with respect to = 0 (i.e., 1 wk after preparing the pots with the seedlings) concentration was significantly higher in inoculated than in noninoculated (NI) pots: The highest DRO losses were found in A D+PGP pots (close to 15-20% higher than NI) and in B D pots (close to 10% higher). The water-extractable DRO fraction was significantly higher at = 30 d (15-25%) compared with = 0 (<5%), probably due to the effects of plant root exudates and biosurfactants produced by the degrader strain. The results of this experiment reflect the importance of the partnerships between plants and bacterial inoculants and demonstrate the relevance of the effect of bacterial biosurfactants and plant root exudates on contaminant bioavailability, a key factor for enhancing diesel rhizodegradation. The association of lupine with D and PGP strains resulted in a promising combination for application in the rhizoremediation of soils with moderate diesel contamination.

Influence of Plant Root Exudates on the Mobility of Fuel Volatile Compounds in Contaminated Soils

Vegetation and its associated microorganisms play an important role in the behaviour of soil contaminants. One of the most important elements is root exudation, since it can affect the mobility, and therefore, the bioavailability of soil contaminants. In this study, we evaluated the influence of root exudates on the mobility of fuel derived compounds in contaminated soils. Samples of humic acid, montmorillonite, and an A horizon from an alumi-umbric Cambisol were contaminated with volatile contaminants present in fuel: oxygenates (MTBE and ETBE) and monoaromatic compounds (benzene, toluene, ethylbenzene and xylene). Natural root exudates obtained from Holcus lanatus and Cytisus striatus and ten artificial exudates (components frequently found in natural exudates) were added to the samples, individually and as a mixture, to evaluate their effects on contaminant mobility. Fuel compounds were analyzed by headspace-gas chromatography-mass spectrometry. In general, the addition of natural and artificial exudates increased the mobility of all contaminants in humic acid. In A horizon and montmorillonite, natural or artificial exudates (as a mixture) decreased the contaminant mobility. However, artificial exudates individually had different effects: carboxylic components increased and phenolic components decreased the contaminant mobility. These results established a base for developing and improving phytoremediation processes of fuel-contaminated soils.

Use of plant growth promoting bacterial strains to improve Cytisus striatus and Lupinus luteus development for potential application in phytoremediation

Plant growth promoting (PGP) bacterial strains possess different mechanisms to improve plant development under common environmental stresses, and are therefore often used as inoculants in soil phytoremediation processes. The aims of the present work were to study the effects of a collection of plant growth promoting bacterial strains on plant development, antioxidant enzyme activities and nutritional status of Cytisus striatus and/or Lupinus luteus plants a) growing in perlite under non-stress conditions and b) growing in diesel-contaminated soil. For this, two greenhouse experiments were designed. Firstly, C. striatus and L. luteus plants were grown from seeds in perlite, and periodically inoculated with 6 PGP strains, either individually or in pairs. Secondly, L. luteus seedlings were grown in soil samples of the A and B horizons of a Cambisol contaminated with 1.25% (w/w) of diesel and inoculated with best PGP inoculant selected from the first experiment. The results indicated that the PGP strains tested in perlite significantly improved plant growth. Combination treatments provoked better growth of L. luteus than the respective individual strains, while individual inoculation treatments were more effective for C. striatus. L. luteus growth in diesel-contaminated soil was significantly improved in the presence of PGP strains, presenting a 2-fold or higher increase in plant biomass. Inoculants did not provoke significant changes in plant nutritional status, with the exception of a subset of siderophore-producing and P-solubilising bacterial strains that resulted in significantly modification of Fe or P concentrations in leaf tissues. Inoculants did not cause significant changes in enzyme activities in perlite experiments, however they significantly reduced oxidative stress in contaminated soils suggesting an improvement in plant tolerance to diesel. Some strains were applied to non-host plants, indicating a non-specific performance of their plant growth promotion. The use of PGP strains in phytoremediation may help plants to overcome contaminant and other soil stresses, increasing phytoremediation efficiency.

Leachability of volatile fuel compounds from contaminated soils and the effect of plant exudates: A comparison of column and batch leaching tests

Volatile fuel compounds such as fuel oxygenates (FO) (MTBE and ETBE) and BTEX (benzene, toluene, ethylbenzene and xylene) are some of the most soluble components of fuel. Characterizing the leaching potential of these compounds is essential for predicting their mobility through the soil profile and assessing the risk of groundwater contamination. Plant root exudates can play an important role in the modification of contaminant mobility in soil-plant systems, and such effects should also be considered in leaching studies. Artificially spiked samples of A and B horizons from an alumi-umbric Cambisol were leached in packed-columns and batch experiments using Milli-Q water and plant root exudates as leaching agents. The leaching potential and rate were strongly influenced by soil-contaminant interactions and by the presence of root exudates. Organic matter in A horizon preferably sorbed the most non-polar contaminants, lowering their leaching potential, and this effect was enhanced by the presence of root exudates. On the other hand, the inorganic components of the B horizon, showed a greater affinity for polar molecules, and the presence of root exudates enhanced the desorption of the contaminants. Column experiments resulted in a more realistic protocol than batch tests for predicting the leaching potential of volatile organic compounds in dissimilar soils.

Phytotoxicity of fuel to crop plants: influence of soil properties, fuel type, and plant tolerance.

The aim of the present study was to determine the effect of fuel-contaminated soils on the germination, survival, and early growth of six crop plants, viz. Brassica oleracea, Trifolium repens, Lactuca sativa, Avena sativa, Pisum sativum, and Zea mays, grown on Cambisol A and B horizons contaminated with gasoline and diesel (0%, 1.25%, 2.5%, 5%, and 10%, w/w). Fuel toxicity was more evident in the B horizon than in the A horizon, and diesel was more toxic than gasoline, probably due to the higher evaporation rate of the latter. Fuels affected the germination and survival of small-seeded plants to a higher extent, reflecting the importance of the seed coat and nutrient reserves for successful plant development in fuel-contaminated soils. In general, root growth was more strongly affected than shoot growth, and plant biomass was more strongly affected than elongation, leading to a less plant branching in the presence of fuel. The findings of this study can be useful for selecting the least fuel-tolerant species as soil contamination bioindicator and for determining the risks of fuel contamination. Due to the low residence time of gasoline components in soil, this phytotoxicity test resulted in an unsuitable bioassay to assess gasoline toxicity.

Draft Genome Sequence of Arthrobacter sp. Strain SPG23, a Hydrocarbon-Degrading and Plant Growth-Promoting Soil Bacterium

ABSTRACT We report here the 4.7-Mb draft genome of Arthrobacter sp. SPG23, a hydrocarbonoclastic Gram-positive bacterium belonging to the Actinobacteria, isolated from diesel-contaminated soil at the Ford Motor Company site in Genk, Belgium. Strain SPG23 is a potent plant growth promoter useful for diesel fuel remediation applications based on plant-bacterium associations.

Characterization and degradation potential of diesel-degrading bacterial strains for application in bioremediation

ABSTRACT Bioremediation of polluted soils is a promising technique with low environmental impact, which uses soil organisms to degrade soil contaminants. In this study, 19 bacterial strains isolated from a diesel-contaminated soil were screened for their diesel-degrading potential, biosurfactant (BS) production, and biofilm formation abilities, all desirable characteristics when selecting strains for re-inoculation into hydrocarbon-contaminated soils. Diesel-degradation rates were determined in vitro in minimal medium with diesel as the sole carbon source. The capacity to degrade diesel range organics (DROs) of strains SPG23 (Arthobacter sp.) and PF1 (Acinetobacter oleivorans) reached 17–26% of total DROs after 10 days, and 90% for strain GK2 (Acinetobacter calcoaceticus). The amount and rate of alkane degradation decreased significantly with increasing carbon number for strains SPG23 and PF1. Strain GK2, which produced BSs and biofilms, exhibited a greater extent, and faster rate of alkane degradation compared to SPG23 and PF1. Based on the outcomes of degradation experiments, in addition to BS production, biofilm formation capacities, and previous genome characterizations, strain GK2 is a promising candidate for microbial-assisted phytoremediation of diesel-contaminated soils. These results are of particular interest to select suitable strains for bioremediation, not only presenting high diesel-degradation rates, but also other characteristics which could improve rhizosphere colonization.

Diesel-Range Organics Extraction and Determination in Environmental Samples by Gas Chromatography‒Mass Spectrometry: Headspace Solid Phase Microextraction vs. Solvent Extraction

An accurate and sensitive analytical method for the determination of diesel-range organics (DRO) is the basis to monitoring and soil remediation studies. In the present work, the determination of DRO in different water and soil samples was optimized. Solvent extraction procedures, i.e. ultrasonic assisted extraction (USAE) (for water samples) and accelerated solvent extraction (ASE) (for soil samples), and a solvent-free procedure, headspace solid phase microextraction (HS-SPME), were optimized to achieve the highest recoveries for the simultaneous determination of all DRO. One hour of USAE for water samples and ASE of soil samples at 100°C, 2000 psi and two extraction cycles lead to analytical recoveries of 70‒100%. Using HS-SPME, 30 min of incubation at 90°C were sufficient to achieve analytical recoveries up to 90% for water and soil samples. HS-SPME enables higher preconcentration factors, which makes this method more appropriate for samples with trace DRO concentrations.

Emergent Properties of Microbial Activity in Heterogeneous Soil Microenvironments: Different Research Approaches Are Slowly Converging, Yet Major Challenges Remain

Over the last 60 years, soil microbiologists have accumulated a wealth of experimental data showing that the bulk, macroscopic parameters (e.g., granulometry, pH, soil organic matter, and biomass contents) commonly used to characterize soils provide insufficient information to describe quantitatively the activity of soil microorganisms and some of its outcomes, like the emission of greenhouse gasses. Clearly, new, more appropriate macroscopic parameters are needed, which reflect better the spatial heterogeneity of soils at the microscale (i.e., the pore scale) that is commensurate with the habitat of many microorganisms. For a long time, spectroscopic and microscopic tools were lacking to quantify processes at that scale, but major technological advances over the last 15 years have made suitable equipment available to researchers. In this context, the objective of the present article is to review progress achieved to date in the significant research program that has ensued. This program can be rationalized as a sequence of steps, namely the quantification and modeling of the physical-, (bio)chemical-, and microbiological properties of soils, the integration of these different perspectives into a unified theory, its upscaling to the macroscopic scale, and, eventually, the development of new approaches to measure macroscopic soil characteristics. At this stage, significant progress has been achieved on the physical front, and to a lesser extent on the (bio)chemical one as well, both in terms of experiments and modeling. With regard to the microbial aspects, although a lot of work has been devoted to the modeling of bacterial and fungal activity in soils at the pore scale, the appropriateness of model assumptions cannot be readily assessed because of the scarcity of relevant experimental data. For significant progress to be made, it is crucial to make sure that research on the microbial components of soil systems does not keep lagging behind the work on the physical and (bio)chemical characteristics. Concerning the subsequent steps in the program, very little integration of the various disciplinary perspectives has occurred so far, and, as a result, researchers have not yet been able to tackle the scaling up to the macroscopic level. Many challenges, some of them daunting, remain on the path ahead. Fortunately, a number of these challenges may be resolved by brand new measuring equipment that will become commercially available in the very near future.

Draft Genome Sequence of Pantoea ananatis GB1, a Plant-Growth-Promoting Hydrocarbonoclastic Root Endophyte, Isolated at a Diesel Fuel Phytoremediation Site Planted with Populus

ABSTRACT We report the 4.76-Mb draft genome of Pantoea ananatis GB1, a Gram-negative bacterium of the family Enterobacteriaceae, isolated from the roots of poplars planted for phytoremediation of a diesel-contaminated plume at the Ford Motor Company site in Genk, Belgium. Strain GB1 promotes plant growth in various hosts and metabolizes hydrocarbons.