María T. Gómez-Sagasti

Microbial Monitoring of the Recovery of Soil Quality During Heavy Metal Phytoremediation

Soil pollution with heavy metals is a worldwide environmental problem. Phytoremediation through phytoextraction and phytostabilization appears to be a promising technology for the remediation of polluted soils. It is important to strongly emphasize that the ultimate goal of a heavy metal remediation process must be not only to remove the heavy metals from the soil (or instead to reduce their bioavailability and mobility) but also to restore soil quality. Soil quality is defined as the capacity of a given soil to perform its functions. Soil microbial properties are increasingly being used as biological indicators of soil quality due to their quick response, high sensitivity, and, above all, capacity to provide information that integrates many environmental factors. Indeed, microbial properties are among the most ecologically relevant indicators of soil quality. Consequently, microbial monitoring of the recovery of soil quality is often carried out during heavy metal phytoremediation processes. However, soil microbial properties are highly context dependent and difficult to interpret. For a better interpretation of microbial properties as indicators of soil quality, they may be grouped within categories of higher ecological relevance, such as soil functions, ecosystem health attributes, and ecosystem services.

PGPRs and nitrogen-fixing legumes: a perfect team for efficient Cd phytoremediation?

Cadmium (Cd) is a toxic, biologically non-essential and highly mobile metal that has become an increasingly important environmental hazard to both wildlife and humans. In contrast to conventional remediation technologies, phytoremediation based on legume–rhizobia symbiosis has emerged as an inexpensive decontamination alternative which also revitalize contaminated soils due to the role of legumes in nitrogen cycling. In recent years, there is a growing interest in understanding symbiotic legume–rhizobia relationship and its interactions with Cd. The aim of the present review is to provide a comprehensive picture of the main effects of Cd in N2-fixing leguminous plants and the benefits of exploiting this symbiosis together with plant growth promoting rhizobacteria to boost an efficient reclamation of Cd-contaminated soils.

Chemical Stabilization of Metal-Contaminated Mine Soil: Early Short-Term Soil-Amendment Interactions and Their Effects on Biological and Chemical Parameters

Chemical stabilization is a cost-effective, environmentally friendly, in situ remediation technology based on the application of organic and/or inorganic amendments to reduce soil metal bioavailability. Our objective was to assess the early short-term effects of organic amendments (sheep manure—SHEEP, poultry litter—POULTRY, cow slurry—COW, paper mill sludge mixed with poultry litter—PAPER), in sterilized and non-sterilized form, on the microbial and chemical properties, as well as on the phytotoxicity, of a Cd, Pb and Zn contaminated soil. Our results provide useful information regarding (1) the effectiveness of amendments for chemical stabilization of mine soil and (2) the impact of microbial populations present in the amendments on soil native microbial communities. Microbial populations present in the amendments did not substantially modify soil microbial functional diversity, as reflected by Biolog EcoPlates™ data, except for PAPER-amended soils. We observed a good correlation between lettuce root elongation (phytotoxicity bioassay) and Cd, Pb, and Zn CaCl2-extractable concentrations in soil. SHEEP and PAPER amendments were particularly effective at increasing soil pH and reducing metal bioavailability and phytotoxicity, while POULTRY and COW led to higher values of soil microbial properties (respiration and functional diversity). Beneficial effects observed under POULTRY at the beginning of the experiment, due to the presence of easily degradable organic matter, were partially lost over time. Our results emphasize the importance of the early monitoring of soil properties (microbial and chemical) and phytotoxicity to properly identify bottlenecks during amendment selection for chemical stabilization, in terms of reduction in metal bioavailability and improvement in soil health.

Agro-industrial wastes as effective amendments for ecotoxicity reduction and soil health improvement in aided phytostabilization

Aided phytostabilization is a technology that uses metal tolerant plants and organic and/or inorganic amendments to reduce soil metal bioavailability, while improving soil health. Our objective was to determine the effects of the application of amendments [sheep manure (SHEEP), poultry litter (POULTRY), cow slurry (COW), and paper mill sludge mixed with poultry litter (PAPER)], together with the growth of a metallicolous Festuca rubra L. population, on (i) chemical and microbial indicators of soil health and (ii) soil ecotoxicity, during the aided phytostabilization of a Zn/Pb contaminated mine soil. Amendment application led to an increase in soil pH, organic matter content, and inorganic salts, resulting in a decrease in Pb and Zn CaCl2-extractable concentrations in soil, which, in turn, contributed to lower ecotoxicity and a stimulation of plant growth and soil microbial communities. The factor most affecting the metal extractability was probably soil pH. POULTRY was the best amendment in terms of increasing plant growth, chlorophylls content, and soil microbial biomass and activity, but resulted in higher levels of phytoavailable Pb and Zn. SHEEP and PAPER were more effective at reducing metal CaCl2-extractability and, consequently, led to lower values of metal accumulation in plant tissues, thereby reducing the risk of metals entering into the food chain. When combined with the application of organic amendments, the metallicolous F. rubra population studied here appears an excellent candidate for aided phytostabilization. Our results indicate that the application of organic amendments is essential for the short-term recovery of highly contaminated metalliferous soils during aided phytostabilization.

Brassica napus has a key role in the recovery of the health of soils contaminated with metals and diesel by rhizoremediation

Contaminated soils are frequently characterized by the simultaneous presence of organic and inorganic contaminants, as well as a poor biological and nutritional status. Rhizoremediation, the combined use of phytoremediation and bioremediation, has been proposed as a Gentle Remediation Option to rehabilitate multi-contaminated soils. Recently, newer techniques, such as the application of metallic nanoparticles, are being deployed in an attempt to improve traditional remediation options. In order to implement a phytomanagement strategy on calcareous alkaline peri-urban soils simultaneously contaminated with several metals and diesel, we evaluated the effectiveness of Brassica napus L., a profitable crop species, assisted with organic amendment and zero-valent iron nanoparticles (nZVI). A two-month phytotron experiment was carried out using two soils, i.e. amended and unamended with organic matter. Soils were artificially contaminated with Zn, Cu and Cd (1500, 500 and 50mgkg-1, respectively) and diesel (6000mgkg-1). After one month of stabilization, soils were treated with nZVI and/or planted with B. napus. The experiment was conducted with 16 treatments resulting from the combination of the following factors: amended/unamended, contaminated/non-contaminated, planted/unplanted and nZVI/no-nZVI. Soil physicochemical characteristics and biological indicators (plant performance and soil microbial properties) were determined at several time points along the experiment. Carbonate content of soils was the crucial factor for metal immobilization and, concomitantly, reduction of metal toxicity. Organic amendment was essential to promote diesel degradation and to improve the health and biomass of B. napus. Soil microorganisms degraded preferably diesel hydrocarbons of biological origin (biodiesel). Plants had a remarkable positive impact on the activity and functional diversity of soil microbial communities. The nZVI were ineffective as soil remediation tools, but did not cause any toxicity. We concluded that rhizoremediation with B. napus combined with an organic amendment is promising for the phytomanagement of calcareous soils with mixed (metals and diesel) contamination.

Early gene expression in Pseudomonas fluorescens exposed to a polymetallic solution

The molecular response of Pseudomonas fluorescens cells exposed to a mixture of heavy metals remains largely unknown. Here, we studied the temporal changes in the early gene expression of P. fluorescens cells exposed to three doses of a polymetallic solution over two exposure times, through the application of a customized cDNA microarray. At the lowest metal dose (MD/4), we observed a repression of the Hsp70 chaperone system, MATE and MFS transporters, TonB membrane transporter and histidine kinases, together with an overexpression of metal transport (ChaC, CopC), chemotaxis and glutamine synthetase genes. At the intermediate metal dose (MD), several amino acid transporters, a response regulator (CheY), a TonB-dependent receptor and the mutT DNA repair gene were repressed; by contrast, an overexpression of genes associated with the antioxidative stress system and the transport of chelates and sulfur was observed. Finally, at the highest metal dose (4MD), a repression of genes encoding metal ion transporters, drug resistance and alginate biosynthesis was found, together with an overexpression of genes encoding antioxidative proteins, membrane transporters, ribosomal proteins, chaperones and proteases. It was concluded that P. fluorescens cells showed, over exposure time, a highly complex molecular response when exposed to a polymetallic solution, involving mechanisms related with chemotaxis, signal transmission, membrane transport, cellular redox state, and the regulation of transcription and ribosomal activity.

Photoprotective Strategies of Mediterranean Plants in Relation to Morphological Traits and Natural Environmental Pressure: A Meta-Analytical Approach

Despite being a small geographic extension, Mediterranean Basin is characterized by an exceptional plant biodiversity. Adaptive responses of this biocoenosis are delineated by an unusual temporal dissociation along the year between optimal temperature for growth and water availability. This fact generates the combination of two environmental stress factors: a period of summer drought, variable in length and intensity, and the occurrence of mild to cold winters. Both abiotic factors, trigger the generation of (photo)oxidative stress and plants orchestrate an arsenal of structural, physiological, biochemical, and molecular mechanisms to withstand such environmental injuries. In the last two decades an important effort has been made to characterize the adaptive morphological and ecophysiological traits behind plant survival strategies with an eye to predict how they will respond to future climatic changes. In the present work, we have compiled data from 89 studies following a meta-analytical approach with the aim of assessing the composition and plasticity of photosynthetic pigments and low-molecular-weight antioxidants (tocopherols, glutathione, and ascorbic acid) of wild Mediterranean plant species. The influence of internal plant and leaf factors on such composition together with the stress responsiveness, were also analyzed. This approach enabled to obtain data from 73 species of the Mediterranean flora, with the genus Quercus being the most frequently studied. Main highlights of present analysis are: (i) sort of photoprotective mechanisms do not differ between Mediterranean plants and other floras but they show higher plasticity indexes; (ii) α−tocopherol among the antioxidants and violaxanthin-cycle pigments show the highest responsiveness to environmental factors; (iii) both winter and drought stresses induce overnight retention of de-epoxidised violaxanthin-cycle pigments; (iv) this retention correlates with depressions of Fv/Fm; and (v) contrary to what could be expected, mature leaves showed higher accumulation of hydrophilic antioxidants than young leaves, and sclerophyllous leaves higher biochemical photoprotective demand than membranous leaves. In a global climatic change scenario, the plasticity of their photoprotective mechanisms will likely benefit Mediterranean species against oceanic ones. Nevertheless, deep research of ecoregions other than the Mediterranean Basin will be needed to fully understand photoprotection strategies of this extremely biodiverse floristic biome: the Mediterranean ecosystem.

cDNA microarray assessment of early gene expression profiles in Escherichia coli cells exposed to a mixture of heavy metals

Many contaminated sites are characterized by the presence of different metals, thus increasing the complexity of toxic responses in exposed organisms. Within toxicogenomics, transcriptomics can be approached through the use of microarrays aimed at producing a genetic fingerprint for the response of model organisms to the presence of chemicals. We studied temporal changes in the early gene expression profiles of Escherichia coli cells exposed to three metal doses of a polymetallic solution over three exposure times, through the application of cDNA microarray technology. In the absence of metals, many genes belonging to a variety of cellular functions were up- and down-regulated over time. At the lowest metal dose, an activation of metal-specific transporters (Cus and ZraP proteins) and a mobilization of glutathione transporters involved in metal sequestration and trafficking was observed over time; this metal dose resulted in the generation of ROS capable of stimulating the transcription of Mn-superoxide dismutase, the assembly of Fe-S clusters and the synthesis of cysteine. At the intermediate dose, an overexpression of ROS scavengers (AhpF, KatG, and YaaA) and heat shock proteins (ClpP, HslV, DnaK, and IbpAB) was observed. Finally, at the highest dose, E. coli cells showed a repression of genes related with DNA mutation correctors (MutY glycopeptidases).

Effectiveness and ecotoxicity of zero-valent iron nanoparticles during rhizoremediation of soil contaminated with Zn, Cu, Cd and diesel

The remediation of soils simultaneously contaminated with organic and inorganic compounds is still a challenging task. The application of metallic nanoparticles, such as zero-valent iron nanoparticles (nZVI), for soil remediation is highly promising, but their effectiveness and potential ecotoxicity must be further investigated. In addition, the performance of nZVI when combined with other remediation strategies is a topic of great interest. Here, we present data on soil chemical (pseudo-total and CaCl2-extractable metal concentrations; petroleum hydrocarbon concentrations) and biological properties (microbial properties and phytotoxicity) after the application of nZVI to soil simultaneously contaminated with Zn, Cu, Cd and diesel, in the absence and presence of other remediation treatments such as the application of an organic amendment and the growth of Brassica napus plants. Soils were artificially contaminated with the abovementioned contaminants. Then, after an aging period of one month, nZVI were applied to the soil and, subsequently, B. napus seeds were sown. Plants were left to grow for one month. Soil samples were collected immediately after artificially contaminating the soil (T1), at sowing (T2) and at harvesting (T3). Overall, the application of nZVI had no effect on contaminant removal, nor on soil microbial parameters. In contrast, it did cause an indirect toxic effect on plant root elongation due to the interaction of nZVI with soil organic matter. These data are useful for researchers and companies interested in the effectiveness and ecotoxicity of zero-valent iron nanoparticles during the remediation of soil contaminated with metals and hydrocarbons, especially when combined with Gentle Remediation Options.

The impact of nanoscale zero-valent iron particles on soil microbial communities is soil dependent

The application of nanoscale zero-valent iron particles (nZVI) for the remediation of contaminated sites is very promising. However, information concerning the ecotoxicity of nZVI on soil microbial communities and, hence, soil quality, is still scarce. We carried out a three-month experiment to evaluate the impact of the application of different concentrations of nZVI (from 1 to 20 mg g DW soil-1) on soil microbial properties in a clay-loam versus a sandy-loam soil. Data on microbial biomass (total bacteria and fungi by qPCR, microbial biomass carbon), activity (β-glucosidase, arylsulphatase and urease activities), and functional (Biolog Ecoplates™) and structural (ARISA, 16S rRNA amplicon sequencing) diversity evidenced that the sandy-loam soil was more vulnerable to the presence of nZVI than the clay-loam soil. In the sandy-loam soil, arylsulphatase activity and bacterial abundance, richness and diversity were susceptible to the presence of nZVI. The high content of clay and organic matter present in the clay-loam soil may explain the observed negligible effects of nZVI on soil microbial properties. It was concluded that the impact of nZVI on soil microbial communities and, hence, soil quality, is soil dependent.