Ángeles Prieto-Fernández

Soil microbial and extractable C and N after wildfire

Abstract The effect of wildfire on soil microbes and extractable C (Cext) and N (Next) changed with respect to the time from burning and soil depth. Initially, microbial biomass C (Cmic) and N (Nmic) were drastically reduced in the soil surface layer (0–5 cm) and reduced by 50% in the subsurface (5–10 cm), whereas Cext increased by 62% in the surface layer and did not significantly change in the subsurface. These parameters were affected for the following 4 years, during which the average reductions in the soil surface and subsurface layers were, respectively, 60% and 50% for Cmic, 70% and 45% for Nmic, 60% and 40% for the ratio Cmic: organic C (Corg) and 70% and 30% for the ratio Nmic: total N (Ntot), while for Cext the surface layer was the only zone consistently affected and Cext decreased by up to 59%. Immediately after a fire, the Cext : Corg ratio increased by 3.5-fold and 2-fold in the surface and subsurface layers, respectively; thereafter for 2 years, it decreased in the surface layer (by up to 45%) while the effect on the subsurface layer was not consistent. The effect of burning on Next lasted 1 year, in which Next increased by up to 7- and 3-fold in the surface and subsurface layers, respectively, while the average Next : Ntot ratio doubled in the surface layer and increased by 34% in the subsurface. During the time in which each parameter was affected by burning, the soil factor explained a high percentage of variance in the fluctuations of Cmic, Nmic, Cmic : Corg and Nmic : Ntot, while those of Next and Next : Ntot, but not those of Cext and Cext : Corg depended on both the soil and its depth. In the burned soils similar patterns of response were found between the following parameters listed in pairs: Cmic and Nmic; Cmic : Corg and Nmic : Ntot; Cext and Next; and Cext : Corg and Next : Ntot. However, after the fire relationships found previously between the parameters studied and many other soils properties were either no longer evident, or were inverted. Although the addition of cellulose to the burned soil favoured fungal mycelium development and increased Cmic and Cext contents, the negative effect of burning on the microbial biomass and the Cext was not counteracted even under incubation conditions suitable for both microbial growth and C mineralization.

Short-term effects of a wildfire on the nitrogen status and its mineralization kinetics in an atlantic forest soil

Abstract A Humic Cambisol developed over granite under Pinus pinaster Sol. located in the Atlantic climate zone, which had been affected by a high intensity wildfire, was studied 1 month after burning. The soil had a very rich organic matter A horizon, 30 cm deep. The effects of the fire on the N status and N mineralization capacity were estimated comparing the surface (0–5 cm) and subsurface (5–10 cm) layers from the burnt soil with the corresponding layers from the same unbumt soil. N mineralization kinetics were determined by aerobic incubation at 28°C for 11 weeks. The fire increased the total N content in the surface layer but not in the subsurface. Total inorganic N, which was mainly in the form of NH + 4 -N, increased after the burning in both layers, whereas NO − 3 -N content, which was very low, only increased in the subsurface layer. The fire increased the N mineralization capacity, but did not modify organic N mineralization behaviour. Ammonification largely predominated over nitrification in both the unburnt and the burnt soils. N mineralization kinetics followed the first order equation N m = N 0 ((1 − e −kt ) but the fire affected the kinetic parameters. The potentially-mineralizable N decreased and the kinetic constant increased in the burnt samples showing that the wildfire reduced the mineralizable organic-N reserves and increased the mineralization rate, thus predicting a rapid depletion of the labile organic N. The temporary ability of the burnt soil to supply available N is recommended to be used to grow an early crop to avoid physical soil degradation.

Pseudometallophytes colonising Pb/Zn mine tailings: A description of the plant–microorganism–rhizosphere soil system and isolation of metal-tolerant bacteria

The plant-microorganism-soil system of three pseudometallophytes (Betula celtiberica, Cytisus scoparius and Festuca rubra) growing in a Pb/Zn mine was characterised. Plant metal accumulation, soil metal fractions (rhizosphere and non-vegetated) and bacterial densities were determined. Total Cd, Pb and Zn in non-vegetated soils was up to 50, 3000 and 20,000 mg kg(-1) dry weight, respectively. The residual fraction dominated non-vegetated soils, whereas plant-available fractions became important in rhizosphere soils. All plant species effectively excluded metals from the shoot. F. rubra presented a shoot:root transport factor of ≤0.2 and this population could be useful in future phytostabilisation trials. Culturable bacterial densities and diversity were low (predominantly Actinobacteria). Rhizosphere soils hosted higher total and metal-tolerant bacterial densities. Seventy-four metal-tolerant rhizobacteria were isolated, and characterised genotypically (BOX-PCR, 16S rDNA) and phenotypically [Cd/Zn tolerance, biosurfactant production and plant growth promoting (PGP) traits]. Several isolates resisted high concentrations of Cd and Zn, and only a few presented PGP traits. Fourteen isolates were evaluated for promoting plant growth of two species (Salix viminalis and Festuca pratensis). Thirteen inoculants enhanced growth of F. pratensis, while only three enhanced growth of S. viminalis. Growth enhancement could not always be related to isolate PGP traits. In conclusion, some isolates show potential application in phytostabilisation or phytoextraction techniques.

Endophytic and rhizoplane bacteria associated with Cytisus striatus growing on hexachlorocyclohexane-contaminated soil: isolation and characterisation

Inoculation of plants with their associated microorganisms is a promising strategy for improving phytoremediation of organic contaminants. Isolation and characterisation of these organisms from plants growing in contaminated sites will permit the identification of candidate strains for re-inoculation studies. The diversity of culturable endophytic and rhizoplane bacteria found in association with Cytisus striatus plants growing at a hexachlorocyclohexane (HCH)-contaminated site was studied. A total of 97 strains of endophytic bacteria were isolated from the root, stem and leaf tissues, and 49 from the rhizoplane. They were further characterised genotypically (BOX-PCR, 16S rDNA sequencing, presence of linA and linB genes) and phenotypically (trace metal tolerance, capacity to produce biosurfactants and plant growth promoting (PGP) traits). Proteobacteria and Actinobacteria dominated the isolate collection, and taxonomic diversity was strongly tissue-specific. The linA and linB genes were not detected in the isolate collection. The majority of isolates had at least one of the PGP traits tested, whereas biosurfactant-producing strains were less frequent. Resistance to more than one trace metal was generally restricted to endophytes isolated from shoot tissues. The PGP characteristics found in an important number of the bacterial isolates obtained in this study could be particularly useful for exploiting the phytoremediation potential of C. striatus.

Rhizobacterial inoculants can improve nickel phytoextraction by the hyperaccumulator Alyssum pintodasilvae

AimRhizobacteria can influence plant growth and metal accumulation. The aim of this study was to evaluate the effect of rhizobacterial inoculants on the Ni phytoextraction efficiency of the Ni-hyperaccumulator Alyssum pintodasilvae.MethodIn a preliminary screening 15 metal-tolerant bacterial strains were tested for their plant growth promoting (PGP) capacity or effect on Ni bioaccumulation. Strains were selected for their Ni tolerance, plant growth promoting traits and Ni solubilizing capacity. In a re-inoculation experiment five of the previously screened bacterial isolates were used to inoculate A. pintodasilvae in two contrasting Ni-rich soils (a serpentine (SP) soil and a sewage sludge-affected agricultural (LF) soil).ResultsPlant growth was greater in serpentine soil (where it grows naturally) than in the LF soil, probably due to Cd phytotoxicity. Rhizobacterial inoculants influenced plant growth and Ni uptake and accumulation, but the effect of the strains was dependent upon soil type. The increase in plant biomass and/or Ni accumulation significantly promoted shoot Ni removal.ConclusionOne strain (Arthrobacter nicotinovorans SA40) was able to promote plant growth and phytoextraction of Ni in both soil types and could be a useful candidate for future field-based trials.

Organic amendments for improving biomass production and metal yield of Ni-hyperaccumulating plants.

Ni phytomining is a promising technology for Ni recovery from low-grade ores such as ultramafic soils. Metal-hyperaccumulators are good candidates for phytomining due to their extraordinary capacity for Ni accumulation. However, many of these plants produce a low biomass, which makes the use of agronomic techniques for improving their growth necessary. In this study, the Ni hyperaccumulators Alyssum serpyllifolium ssp. lusitanicum, A. serpyllifolium ssp. malacitanum, Alyssum bertolonii and Noccaea goesingense were evaluated for their Ni phytoextraction efficiency from a Ni-rich serpentine soil. Effects of soil inorganic fertilisation (100:100:125kgNPKha(-1)) and soil organic amendment addition (2.5, 5 or 10% compost) on plant growth and Ni accumulation were determined. All soil treatments greatly improved plant growth, but the highest biomass production was generally found after addition of 2.5 or 5% compost (w/w). The most pronounced beneficial effects were observed for N. goesingense. Total Ni phytoextracted from soils was significantly improved using both soil treatments (inorganic and organic), despite the decrease observed in soil Ni availability and shoot Ni concentrations in compost-amended soils. The most promising results were found using intermediate amount of compost, indicating that these types of organic wastes can be incorporated into phytomining systems.

Rhizosphere Microbial Densities and Trace Metal Tolerance of the Nickel Hyperaccumulator Alyssum Serpyllifolium Subsp. Lusitanicum

In this study we determine culturable microbial densities (total heterotrophs, ammonifiers, amylolytics and cellulolytics) and bacterial resistance to Co, Cr, and Ni in bulk and rhizosphere soils of three populations of the Ni-hyperaccumulator Alyssum serpyllifolium subsp. lusitanicum and the excluder Dactylis glomerata from ultramafic sites (two populations in Northeast (NE) Portugal (Samil (S), Morais (M)) and one population in Northwest (NW) Spain (Melide (L)). The relationship between bioavailable metal concentrations (H2O-soluble) and microbial densities were analysed. Significant differences in microbial densities and metal-resistance were observed between the two species and their three populations. The hyperaccumulator showed higher microbial densities (except cellulolytics) and a greater rhizosphere effect, but this was only observed in S and M populations. These populations of A. serpyllifolium also showed selective enrichment of Ni-tolerant bacteria at the rhizosphere where Ni solubility was enhanced (densities of Ni-resistant bacteria were positively correlated with H2O-soluble Ni). These rhizobacteria could solubilise Ni in the soil and potentially improve phytoextraction strategies.

Bacterially Induced Weathering of Ultramafic Rock and Its Implications for Phytoextraction

ABSTRACT The bioavailability of metals in soil is often cited as a limiting factor of phytoextraction (or phytomining). Bacterial metabolites, such as organic acids, siderophores, or biosurfactants, have been shown to mobilize metals, and their use to improve metal extraction has been proposed. In this study, the weathering capacities of, and Ni mobilization by, bacterial strains were evaluated. Minimal medium containing ground ultramafic rock was inoculated with either of two Arthrobacter strains: LA44 (indole acetic acid [IAA] producer) or SBA82 (siderophore producer, PO4 solubilizer, and IAA producer). Trace elements and organic compounds were determined in aliquots taken at different time intervals after inoculation. Trace metal fractionation was carried out on the remaining rock at the end of the experiment. The results suggest that the strains act upon different mineral phases. LA44 is a more efficient Ni mobilizer, apparently solubilizing Ni associated with Mn oxides, and this appeared to be related to oxalate production. SBA82 also leads to release of Ni and Mn, albeit to a much lower extent. In this case, the concurrent mobilization of Fe and Si indicates preferential weathering of Fe oxides and serpentine minerals, possibly related to the siderophore production capacity of the strain. The same bacterial strains were tested in a soil-plant system: the Ni hyperaccumulator Alyssum serpyllifolium subsp. malacitanum was grown in ultramafic soil in a rhizobox system and inoculated with each bacterial strain. At harvest, biomass production and shoot Ni concentrations were higher in plants from inoculated pots than from noninoculated pots. Ni yield was significantly enhanced in plants inoculated with LA44. These results suggest that Ni-mobilizing inoculants could be useful for improving Ni uptake by hyperaccumulator plants.

Nickel Solubilizing Capacity and Characterization of Rhizobacteria Isolated from Hyperaccumulating and Non-Hyperaccumulating Subspecies of Alyssum Serpyllifolium

Bacterial strains were isolated from the rhizosphere of three populations of the Ni-hyperaccumulator Alyssum serpyllifolium subsp. lusitanicum (A. pintodasilvae; M, S, and L), one population of Ni-hyperaccumulator A. serpyllifolium subsp. malacitanum (A. malacitanum; SB), and one population of the non-hyperaccumulator A. serpyllifolium subsp. serpyllifolium (A. serpyllifolium; SN). Isolates were characterized genotypically by BOX-PCR genomic DNA fingerprinting and comparative sequence analysis of partial 16S rRNA gene, and phenotypically by their Ni tolerance (0–10 mM), presence of plant growth promoting traits (indoleacetic acid (IAA)-, siderophore-, or organic acid-production, and phosphate solubilization) or capacity to produce biosurfactants. Among the collection of rhizobacteria, 84 strains were selected (according to their BOX-PCR profiles and phenotypic characteristics) to assess their ability to modify Ni extractability from Ni-rich (serpentine) soils. Metabolites produced by 13 of the isolates mobilized soil Ni (originating from the rhizosphere of both Ni-hyperaccumulators and non-hyperaccumulator). In contrast, Ni extraction using culture medium filtrates which had supported the growth of 29 strains was significantly reduced. The remaining strains had no effect on Ni mobility. Bacterial induced Ni mobilization was not related to Ni resistance or the phenotypic traits tested. Isolates with potential use in phytoremediation techniques will be further studied in a plant-microorganism-soil system.

Exogenous treatments with phytohormones can improve growth and nickel yield of hyperaccumulating plants.

The application of plant growth regulators (PGRs) or phytohormones could be an interesting option for stimulating biomass production of hyperaccumulating plants and, consequently, their metal phytoextraction capacity. The effect of exogenous applications of phytohormones (PGR) on the Ni phytoextraction capacity of four Ni hyperaccumulating species (Alyssum corsicum, Alyssum malacitanum, Alyssum murale and Noccaea goesingense) was evaluated. Four different commercially available phytohormones (B, C, K and P) based on gibberellins, cytokinins and auxins were applied to the plant aerial tissues. Each product was applied at three different concentrations (B1-3, C1-3, K1-3 and P1-3). The effect on biomass production was dependent on the species, the PGR type and the concentration at which it was applied. Two of the four products (K and P) consistently increased biomass production compared to untreated control plants in all four plant species. On the other hand, all four products led to a significant increase in the number of branches (and leaves in the case of N. goesingense) of all four species compared to control plants. Application of phytohormones generally led to a reduction in shoot Ni concentration. Nonetheless, in some cases as a consequence of the increase observed in biomass after the application of phytohormones a significant increase in the Ni phytoextraction efficiency was also observed (but this was species- and PGR type-dependent). The results show that PGRs can be successfully used to improve the growth and biomass production of hyperaccumulating species such as Alyssum and Noccaea. However, an increase in biomass did not always lead to a higher Ni removal, and the most effective PGR for increasing Ni removal was the IAA-based product.