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1.

Purpose

This study aimed to evaluate the effect of combination of alkyl polyglucoside (APG) and nitrilotriacetic acid (NTA) on improving the efficiency of phytoremediation for pyrene and lead (Pb) co-contaminated soil by Scirpus triqueter.

Materials and methods

Seedlings of S. triqueter with a similar size and biomass (3 g/pot) were grown on 2-month aged soil contaminated with 184.5 mg kg?1of pyrene and 454.3 mg kg?1 of Pb at pH?=?8.3. After growth for 10 days, different doses of APG and NTA were added into the soil. After 60 days, the height of plants, Pb concentrations in plants, and pyrene amounts in soil were determined.

Results and discussion

Combined application of NTA and APG with lower dosage (1 + 1 g kg?1 soil and 1 + 2 g kg?1 soil) had no notable negative influence on the growth of S. triqueter. Moreover, significant synergy on Pb accumulation in S. triqueter was achieved with APG and NTA combined application. Besides, the dissipation of pyrene from soil after 60-day planting was increased in APG and NTA treatments when compared with the control treatments. Application of APG alone or combined with NTA had greater effect on enhancing dissipation of pyrene from soil than NTA alone.

Conclusions

This study demonstrated that the remediation of Pb and pyrene co-contaminated soil by S. triqueter can be enhanced by combined application of APG and NTA. Long-term evaluation of this strategy is needed in co-contaminated field sites.
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2.

Purpose

This study delineated the effect of calcium (Ca) and ethylenediaminetetraacetic acid (EDTA) addition on arsenic (As) accumulation and physiological attributes of Vicia faba L.

Materials and methods

Two separate experiments were performed. In the first experiment, V. faba seedlings, grown under hydroponic conditions, were exposed to three levels of As (25, 125 and 250 μM) in the presence and absence of three levels of EDTA (25, 125, 250 μM) and calcium (CaCl2: 1, 5 and 10 mM). The effect of EDTA and Ca on As accumulation and physiological attributes of V. faba was assessed by determining As contents in roots and shoot, chlorophyll contents, H2O2 contents, and lipid peroxidation in young and old leaves. In the second experiment, V. faba seeds were grown in As-contaminated sand culture using the same treatment plan.

Results and discussion

The accumulation and toxicity of As to V. faba plants increased with increasing As levels in nutrient solution. Arsenic exposure enhanced the production of reactive oxygen species (ROS) in both roots and leaves, which resulted in lipid peroxidation and decreased chlorophyll contents. The presence of both EDTA and Ca, in general, significantly decreased As accumulation by V. faba seedlings, Ca being more effective than EDTA. Both the amendments decreased As-induced reactive oxygen species (ROS) production and lipid peroxidation. In the case of chlorophyll contents, EDTA significantly decreased chlorophyll contents, while Ca significantly increased chlorophyll contents compared to As. The effect of all the treatments was more pronounced in roots than leaves and in young leaves compared to old leaves.

Conclusions

It is proposed that EDTA and Ca greatly affect As accumulation and toxicity to V. faba, and the effect varies greatly with their applied levels as well as type and age of plant organs. The germinating seedlings of V. faba may be preferred for risk assessment studies, while transplanting 1-week-old V. faba seedlings to As-contaminated soils can decrease its toxicity.
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3.

Purpose

This study investigated the extent of metal accumulation by plants colonizing a mining area in Yazd Province in Central Iran. It also investigated the suitability of these plants for phytoextraction and phytostabilization as two potential phytoremediation strategies.

Materials and methods

Plants with a high bioconcentration factor (BCF) and low translocation factor (TF) have the potential for phytostabilization, whereas plants with both BCFs and TFs >1 may be appropriate for phytoextraction. In this study, both shoots and roots of 40 plant species and associated soil samples were collected and analyzed for total concentrations of trace elements (Pb, Zn, and Ag). BCFs and TFs were calculated for each element.

Results and discussion

Nonnea persica, Achillea wilhelmsii, Erodium cicutarium, and Mentha longifolia were found to be the most suitable species for phytostabilization of Pb and Zn. Colchicum schimperi, Londesia eriantha, Lallemantia royleana, Bromus tectorum, Hordeum glaucum, and Thuspeinantha persica are the most promising species for element phytoextraction in sites slightly enriched by Ag. Ferula assa-foetida is the most suitable species for phytostabilization of the three studied metals. C. schimperi, L. eriantha, L. royleana, B. tectorum, M. longifolia, and T. persica accumulated Ag, albeit at low level.

Conclusions

Our preliminary study shows that some native plant species growing on this contaminated site may have potential for phytoremediation.
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4.

Purpose

The possibility of using chemical and microbial additives to enhance the phytoextraction of mercury (Hg) and arsenic (As) from a multi-contaminated soil could be very effective, leading to a significant saving in terms of time and costs of the reclamation. The aim of this study was to evaluate the efficacy of the addition of (i) thiosulfate and (ii) metal-tolerant bacteria isolated from the polluted soil having plant growth promotion (PGP) potential to perform As and Hg phytoextraction by Brassica juncea and Lupinus albus.

Materials and methods

A collection of 13 bacterial isolates able to tolerate As and Hg was obtained from the contaminated soil, identified by partial 16S rRNA gene sequencing and tested in vitro for PGP activities. The most promising strains were further tested in vivo for the evaluation of plant growth ability and rhizocompetence on model plants. Pot experiments were conducted in microcosms, with polluted soil vegetated with B. juncea and L. albus. Ammonium thiosulfate and potassium dihydrogen phosphate were used as mobilizing agents, together with a bacterial consortium composed by the most promising PGP isolates.

Results and discussion

Thirteen indigenous metal-tolerant bacterial strains were isolated, and their in vitro characterization highlighted their great potential in assisting the phytoremediation process; most of them tolerated both trace elements and showed, at the same time, multiple PGP traits. The results were confirmed in vivo on model plants and lead to the selection of the most promising PGP strains to be applied in microcosm-scale phytoextraction experiments. Thiosulfate addition significantly increased the mobilization of both elements, promoting bioavailability and phytoextraction. When a selected bacterial consortium was supplemented in addition to thiosulfate, the efficacy of the phytoaccumulation was increased up to 85 % for As and up to 45 % for Hg.

Conclusions

The use of the common fertilizer thiosulfate appeared to have great potential in phytoextraction practices since it was able to facilitate the uptake by plants of both Hg and As. Moreover, the application of a consortium of indigenous PGP bacteria (PGPB) produced a further positive effect on the plant biomass, supporting and enhancing the phytoextraction strategy, thus demonstrating their potential in a microbe-assisted phytoremediation intervention.
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5.

Purpose

The subjects of this study were to investigate the remediating potential of the co-cultivation of Pleurotus eryngii and Coprinus comatus on soil that is co-contaminated with heavy metal (cadmium (Cd)) and organic pollutant (endosulfan), and the effects of the co-cultivated mushrooms on soil biochemical indicators, such as laccase enzyme activity and bacterial counts.

Materials and methods

A pot experiment was conducted to investigate the combined bioremediation effects on co-contaminated soil. After the mature fruiting bodies were harvested from each pot, the biomass of mushrooms was recorded. In addition, bacterial counts and laccase enzyme activity in soil were determined. The content of Cd in mushrooms and soil was detected by the flame atomic absorption spectrometry (FAAS), and the variations of Cd fractions in soil were determined following the modified BCR sequential extraction procedure. Besides, the residual endosulfan in soil was detected by gas chromatography-mass spectrometry (GC-MS).

Results and discussion

The results indicated that co-cultivation of P. eryngii and C. comatus exerted the best remediation effect on the co-contaminated soil. The biomass of mushroom in the co-cultivated group (T group) was 1.57–13.20 and 19.75–56.64% higher than the group individually cultivated with P. eryngii (P group) or C. comatus (C group), respectively. The concentrations of Cd in the fruiting bodies of mushrooms were 1.83–3.06, 1.04–2.28, and 0.67–2.60 mg/kg in T, P, and C groups, respectively. Besides, the removal rates of endosulfan in all treatments exceeded 87%. The best bioremediation effect in T group might be caused by the mutual promotion of these two kinds of mushrooms.

Conclusions

The biomass of mushroom, laccase activity, bacterial counts, and Cd content in mushrooms were significantly enhanced, and the dissipation effect of endosulfan was slightly higher in the co-cultivated group than in the individually cultivated groups. In this study, the effect of co-cultivated macro fungi P. eryngii and C. comatus on the remediation of Cd and endosulfan co-contaminated soil was firstly reported, and the results are important for a better understanding of the co-remediation for co-contaminated soil.
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6.

Purpose

Fruiting vegetables are generally considered to be safer than other vegetables for planting on cadmium (Cd)-contaminated farms. However, the risk of transferring Cd that has accumulated in the stems and leaves of fruiting vegetables is a major issue encountered with the usage of such non-edible parts. The objective of this study was to resolve the contribution of arbuscular mycorrhizal (AM) fungi to the production of low-Cd fruiting vegetables (focusing on the non-edible parts) on Cd-contaminated fields.

Materials and methods

An 8-week pot experiment was conducted to investigate the acquisition and translocation of Cd by cucumber (Cucumis sativus L.) plants on an unsterilized Cd-contaminated (1.6 mg kg?1) soil in response to inoculation with the AM fungus, Funneliformis caledonium (Fc) or Glomus versiforme (Gv). Mycorrhizal colonization rates of cucumber roots were assessed. Dry biomass and Cd and phosphorus (P) concentrations in the cucumber shoots and roots were all measured. Soil pH, EC, total Cd, phytoavailable (DTPA-extractable) Cd, available P, and acid phosphatase activity were also tested.

Results and discussion

Both Fc and Gv significantly increased (P?<?0.05) root mycorrhizal colonization rates and P acquisition efficiencies, and thus the total P acquisition and biomass of cucumber plants, whereas only Fc significantly increased (P?<?0.05) soil acid phosphatase activity and the available P concentration. Both Fc and Gv significantly increased (P?<?0.05) root to shoot P translocation factors, inducing significantly higher (P?<?0.05) shoot P concentrations and shoot/root biomass ratios. In contrast, both Fc and Gv significantly decreased (P?<?0.05) root and shoot Cd concentrations, resulting in significantly increased (P?<?0.05) P/Cd concentration ratios, whereas only Gv significantly decreased (P?<?0.05) the root Cd acquisition efficiency and increased (P?<?0.05) the root to shoot Cd translocation factor. Additionally, AM fungi also tended to decrease soil total and phytoavailable Cd concentrations by elevating plant total Cd acquisition and soil pH, respectively.

Conclusions

Inoculation with AM fungi increased the P acquisition and biomass of cucumber plants, but decreased plant Cd concentrations by reducing the root Cd acquisition efficiency, and resulted in a tendency toward decreases in soil phytoavailable and total Cd concentrations via increases in soil pH and total Cd acquisition by cucumber plants, respectively. These results demonstrate the potential application of AM fungi for the production of fruiting vegetables with non-edible parts that contain low Cd levels on Cd-contaminated soils.
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7.

Purpose

Studying the rate of chelant degradation is important to select environmental friendly compounds to assist phytoextraction. The objective of the present study was to evaluate degradation rate of complexes formed between synthetic or organic chelants and Pb aiming to increase the efficiency of phytoextraction while reducing adverse effects resulting from the Pb leaching.

Materials and methods

The following six chelating agents were tested: citric acid P.A., commercial citric acid, glutamic acid P.A., monosodium glutamate, nitrilotriacetic acid (NTA), and ethylenediaminetetraacetic acid (EDTA), besides a control treatment (no addition of chelating agent); they were applied at a concentration of 10 mmol dm?3 in pots containing 1 dm3 of Pb-contaminated soil.

Results and discussion

The results of this study showed that commercial citric acid adequately solubilized Pb to levels suitable for plant uptake and showed relatively rapid biodegradation in soil. Therefore, this commercial product may be a highly promising alternative for phytoextraction studies in the field. EDTA and NTA demonstrated high Pb solubilization ability but degraded comparatively slowly; therefore, they are not recommended for use in phytoextraction due to environmental risks regarding metal leaching.

Conclusions

The results of this study showed that commercial citric acid adequately solubilized Pb to levels suitable for plant uptake and showed relatively rapid biodegradation in soil, which is associated with a low risk of groundwater contamination. Therefore, this environmental friendly and low-cost product may be a highly promising alternative for inducing Pb phytoextraction.
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8.

Purpose

Applications of mineral and organic fertilizer increased soil cadmium (Cd) and could enhance Cd concentrations in edible crops, respectively. Although red mud (RMD) effectively decreased metal bioavailability in soil, the influence of RMD addition on vegetable growth and metal accumulation under high fertilization conditions has rarely been addressed. The aim of this study was to investigate the effects of raw RMD addition on cole growth, quality, and nutrition and Cd accumulation under high fertilization conditions.

Materials and methods

Pot experiments with cole (Brassica campestris L.) were carried out in a greenhouse. Three treatments, CK (with no mineral fertilizer and RMD addition), CT (more than 2.5 times conventional level of mineral fertilizer applied without any RMD), and RM (more than 2.5 times conventional level of mineral fertilizer applied with RMD added at 0.4 % w/w), were established. After 40 days, the cole plants and soils of every replicate of all treatments were sampled. The Cd, biomass, vitamin C (VC), and total nitrogen and phosphorus of the cole plant samples and the Cd, pH, nitrate, and phosphorus of the soil samples were determined.

Results and discussion

In contrast to the CT treatment, RM treatment did not significantly influence the biomass and nitrate concentration of the aboveground cole. However, it significantly reduced the Cd content in cole shoots and its bioaccumulation factors by 30.0 and 28.5 %, respectively. The reduction of bioavailable Cd in soil by RMD sorption and the competition with calcium released from RMD led to low Cd assimilation by root. Finally, less Cd was transported to aboveground plant parts in the RM treatment compared with the CT treatment. RMD addition markedly enhanced the total nitrogen in cole shoots by >16.0 %, and the VC by 20.9 %. The promotion of bacterial abundance and soil enzyme activity by RMD addition and calcium release from RMD generated substantial plant available nitrogen.

Conclusions

With large rate of mineral fertilizer application to the soil, RMD (0.4 %, w/w) addition did not significantly influence the biomass, nitrate, and VC of aboveground cole; however, it significantly reduced the Cd and markedly enhanced the total nitrogen in cole shoots. This study provides valuable information for the safe application of RMD in vegetable production.
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9.

Purpose

Soils formed in metallic mines and serpentinite quarries, among other unfavourable features, have high levels of heavy metals. They can release into the environment causing surface and subsurface water contamination, uptake by plants, their accumulation in the food chain and adverse effects on living organisms. In this work, we studied the magnitude of the soils’ toxic effects not only on spontaneous plants but also on two species with phytoremediation potential.

Materials and methods

Several soils from two different exploitations were selected: a lead and zinc mine and a serpentinite quarry. Soils were characterized, and the pseudo-total and extractable contents of Co, Cr and Ni in soils from a serpentinite quarry were determined. The Cd, Pb and Zn pseudo-total and extractable contents were determined in soils developed in the Pb/Zn abandoned mine. Using a biotest, the chronic toxicity of the soil samples on higher plants was determined. Festuca ovina L., Cytisus scoparius (L.) Link., Sinapis alba L. and Brassica juncea L. were selected, the first two because they are spontaneous plants in the study areas and the last two because they have heavy metal phytoremediation potential.

Results and discussion

Pseudo-total contents of Co, Cr and Ni in the serpentinite quarry soils and of Zn, Pb and Cd in the Zn/Pb mine soils exceed generic reference levels. CaCl2 is the reactant that extracts the highest proportion of Co, Cr and Ni in the quarry soils and EDTA the largest proportion of Pb Zn and Cd content in the mine soils. The germination index values based on seed germination and root elongation bioassays revealed increasing plant sensitivity to the mine soils in the following order: B. juncea?<?S. alba?<?F. ovina?<?C. scoparius. The wide range of GI values indicates that the response of test plants to soil heavy metals depended on their concentrations and soil characteristics, especially pH and organic matter content.

Conclusions

The pollution index indicates severe Cd, Pb and Zn contamination in the mine soils, as well as high Cr and Ni and moderate Co contamination in the serpentinite quarry soils. The performed biotests were suitable for identifying toxic soils and showed that the studied soils are toxic to the spontaneous plants, more to C. scoparius than to F. ovina. They also indicate that the mine soils are more toxic than the quarry soils for both species.
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10.

Purpose

The in situ stabilization of multielement-contaminated agricultural soils has limited effectiveness when using common single amendments. This study examined the use of drinking water treatment residues (WTR), based on (hydr)oxides of Fe, Al, or Mn, as a cost-effective solution to optimize the immobilization of metals (Cd, Pb, Zn) and As.

Materials and methods

Trace elements (TE) bioavailability was assessed under semi-controlled conditions in a pot study cultivating winter wheat (Triticum aestivum L. cv. Tiger) until maturity. An Fe-based WTR and a Mn-based WTR, applied at rates of 0.5 and 1% (m/m), were related to effects of lime marl (LM) application. Additionally, a bioassay with earthworms (Dendrobaena veneta) was conducted. Both bioassays were compared with measurements of NH4NO3-soluble, diffusive gradients in thin film (DGT)-available and soil solution TE concentrations, representing well-established surrogates for mimicking the bioavailable element fractions in soil.

Results and discussion

The application of the Fe-based WTR reduced As accumulation in vegetative wheat tissues (by up to 75%) and earthworms (by up to 41%), which corresponded with the findings from soil chemical analyses and improved plant growth and earthworm body weight. However, As concentrations in cereal grains were not affected, Cd or Pb accumulation by wheat was not mitigated, and Zn uptake was enhanced. By contrast, the Mn-based WTR effected the greatest reduction in Pb uptake, and lowered Cd transfer to wheat grain (by up to 25%). Neither the NH4NO3-soluble nor DGT-available concentrations matched with Cd and Zn accumulation in plants or earthworms, indicating interferences due to competition for binding sites according to the biotic ligand model.

Conclusions

The results obtained in this study suggest that a bioassay with key species prior to field application should be mandatory when designing in situ stabilization options. The application of WTR to an agricultural soil strongly affected TE bioavailability to plants and earthworms. Low application rates tended to improve biomass production of biota. Higher application rates involved risks (e.g., P fixation, TE inputs), and none of the amendments tested could immobilize all targeted elements.
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11.

Purpose

This study aims to investigate the response of a high biomass producer non-hyperaccumulator legume plant species, Dolichos lablab L., to cadmium (Cd) stress for phytoremediation process.

Materials and methods

Three individual experiments were carried out to assess physiological and biochemical parameters to support the use of this plant species as a phytoremediator. The first experiment was carried out in Cd-contaminated soil while the second and third experiments were conducted in sand in which Cd was applied to study biochemical responses. Analysis of mineral nutrition, phytoremediation parameters, antioxidant response, and protein identification by gel-based proteomics were performed.

Results and discussion

Good tolerance to Cd under moderate level of contamination was observed. Mineral nutrition was little affected, and phytoremediation index was satisfactory. Additionally, biochemical responses based on antioxidant enzyme analysis were well responsive in roots, reflecting the capacity of Cd stress attenuation in this organ. A proteomic analysis revealed positive regulation of root proteins involved in carbohydrate, amino acids, nitrogen metabolism, and abiotic/biotic stress response, which together may contribute to create a scenario to overcome Cd-induced stress.

Conclusions

Based on the physiological and biochemical results, we concluded that D. lablab L. is suitable for phytoremediation/phytostabilization purposes.
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12.

Purpose

This study aimed at investigating the rhizosphere effects of Populus euramericana Dorskamp on the mobility of Zn, Pb and Cd in contaminated technosols from a former smelting site.

Materials and methods

A rhizobox experiment was conducted with poplars, where the plant stem cuttings were grown in contaminated technosols for 2 months under glasshouse conditions. After plant growth, rhizosphere and bulk soil pore water (SPW) were sampled together. SPW properties such as pH, dissolved organic carbon (DOC) and total dissolved concentrations of Zn, Pb and Cd were determined. The concentrations of Zn, Pb and Cd in plant organs were also determined.

Results and discussion

Rhizosphere SPW pH increased for all studied soils by 0.3 to 0.6 units compared to bulk soils. A significant increase was also observed for DOC concentrations regardless of the soil type or total metal concentrations, which might be attributed to the plant root activity. For all studied soils, the rhizosphere SPW metal concentrations decreased significantly after plant growth compared to bulk soils which might be attributed to the increase in pH and effects of root exudates. Zn, Pb and Cd accumulated in plant organs and the higher metal concentrations were found in plant roots compared to plant shoots.

Conclusions

The restricted transfer of the studied metals to the plant shoots confirms the potential role of this species in the immobilization of these metals. Thus, P. euramericana Dorskamp can be used for phytostabilization of technosols.
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13.

Purpose

Nitrogen (N) application in excess of assimilatory capacity for aquaculture ponds can lead to water-quality deterioration through ammonia accumulation with toxicity to fish. Ammonia-oxidizing archaea (AOA) and bacteria (AOB) potentially process extra ammonium, so their abundance and diversity are of great ecological significance. This study aimed to reveal variations in communities of AOA and AOB as affected by aquaculture activities.

Materials and methods

From June to September 2012, water and sediments were sampled monthly in three ponds feeding Mandarin fish in a suburb of Wuhan City, China. Molecular methods based on ammonia monooxygenase (amoA) gene were used to determine abundance and diversity of AOA and AOB in the sediments.

Results and discussion

The pond with the highest fish stock had the highest nutrient loadings in terms of different forms of N and carbon (C) in both sediment and water. The abundance and diversity of AOB were significantly higher than those of AOA in the sediment. The AOB abundance showed a significantly positive relationship to concentration of soluble reactive phosphorus (SRP) in interstitial water, and both abundance and diversity of AOA were significantly negative to concentration of ammonium in interstitial water. Furthermore, AOA species affiliated to Nitrososphaera-like and Nitrosophaera Cluster was distinguishable from those observed in other aquaculture environments.

Conclusions

Nutrients in sediment were enriched by intensive aquaculture activity, among which organic N and C, together with ammonium and SRP, shaped the communities of ammonia oxidizers, with AOB dominating over AOA in terms of abundance and diversity.
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14.

Purpose

Phosphate (P) fertilizers are being widely used to increase crop yield, especially in P-deficient soils. However, repeated applications of P could influence trace element bioaccumulation in crops. The effects of 5-year P enrichment on trace element (Cu, Zn, Cd, Pb, As, and Hg) accumulation in Oryza sativa L. were thus examined.

Materials and methods

Two paddy soils with different initial P availabilities were amended with and without P fertilizer from 2009 to 2013. Trace elements and P levels in rice and soils were analyzed.

Results and discussion

In soil initially with limited P, P amendment enhanced grain Pb, As, and Hg concentrations by 1.8, 1.5, and 1.4-fold, respectively, but tended to decrease the grain Cd level by 0.73-fold, as compared to the control. However, in soil initially with sufficient P, P amendment tended to reduce accumulation of all examined elements in rice grain.

Conclusions

Phosphate amendment in initially P-limited and P-sufficient soils had different effects on trace element availability in soil (as reflected by extractable element) and plant physiology (growth and metal translocation), resulting in contrasting patterns of trace element accumulation in rice between the two types of soils. Our study emphasized the necessity to consider the promoting effects of P on Pb, As, and Hg accumulation in grain in initial P-deprived soil.
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15.

Purpose

Chickpea is generally cultivated after seed treatment with host-specific Mesorhizobium ciceri, the nitrogen-fixing bacterium forming root nodules. Some species of free-living cyanobacteria are capable of nitrogen fixation. We examined the rhizosphere microbiota changes and the potential for plant growth promotion by applying a free-living, nitrogen-fixing cyanobacterium and the biofilm formulation of cyanobacterium with M. ciceri, relative to M. ciceri applied singly, to two each of desi and kabuli varieties of chickpea.

Materials and methods

Denaturing gradient gel electrophoresis (DGGE) profiles of archaeal, bacterial and cyanobacterial communities and those of phospholipid fatty acids (PLFAs) were obtained to evaluate the changes of the microbial communities in the chickpea rhizosphere. Plant growth attributes, including the pod yields and the availabilities of soil macronutrients and micronutrients, were monitored.

Results and discussion

The DGGE profiles showed distinct and characteristic changes due to the microbial inoculation; varietal differences exerted a marked influence on the archaeal and cyanobacterial communities. However, bacterial communities were modulated more by the type of microbial inoculants. Abundance of Gram-negative bacteria (in terms of notional PLFAs) differed between the desi and the kabuli varieties inoculated with M. ciceri alone, and the principal component analysis of PLFA profiles confirmed the characteristic effect of microbial inoculants tested. Microbial inoculation led to increases in the 100-seed weight and differential effects on the concentrations of available nitrogen and phosphorus, and those of iron, zinc and copper, suggesting their increased cycling in the rhizosphere.

Conclusions

Microbial inoculation of chickpea brought out the characteristic changes in rhizosphere microbiota. Consequently, the growth promotion of chickpea and nutrient cycling in its rhizosphere distinctively differed. Further studies are needed to analyse the association and dynamic changes in the microbial communities to define the subset of microorganisms selected by chickpea in its rhizosphere and the influence of microbial inoculation.
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16.

Purpose

Soil metal pollution is a widespread problem around the world and remediation of these soils is difficult. The objective of this study was to investigate the effect of two different strategies on the chemical fractions of metals in a soil of a depleted copper mine: (1) amending with a mixture of a technosol made of wastes and biochar and (2) amending combined with planting vegetation (Brassica juncea).

Materials and methods

A 3-month greenhouse experiment was carried out to evaluate the effects of organic amendments and vegetation on the metal fractionation of Cu, Ni, Pb and Zn in a mine soil of the depleted copper mine at Touro (Spain). We compared the influence of organic amendments alone (technosol?+?biochar) and combined with mustard plants (Brassica juncea L.).

Results and discussion

The results showed that amending with a technosol made of wastes promoted plant growth (from 0.7 to 2.9 g of biomass produced) and reduced the CaCl2-extractable metal concentration in soil, reduced the mobility factor of Cu from 18.3 to 1.6, Ni from 47.5 to 2.3 and Pb from 17.9 to 2.1, and also reduced the concentration of metals in the mobile soil fractions. It was not possible to grow up Brassica juncea plants in the untreated settling pond soil due to the extremely degraded conditions of that soil. However, the application of the used technosol increased the Pb and Zn pseudototal concentrations in the amended soils.

Conclusions

We conclude that the combination of amending with wastes and planting B. juncea provides little additional benefit for remediating a metal-polluted soil compared with incorporation of wastes alone.
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17.

Purpose

The presence of high concentrations of trace elements (TEs) in mine soils like those in the Sierra Minera of La Unión-Cartagena (SE Spain) limits the development of a vegetation cover on such sites, and pollution dispersion by water and wind erosion represents a serious risk for the surrounding ecosystems. The aim of this study was to evaluate different phytostabilisation procedures based on the co-culture of a legume (Bituminaria bituminosa) and a high-biomass (Piptatherum miliaceum) species for this type of soils.

Materials and methods

A pot experiment was carried out where B. bituminosa was tested as a soil pre-treatment strategy. Five different procedures were followed to study the growth stimulation or competition of both species in a contaminated soil from the Sierra Minera: (i) sowing of P. miliaceum without B. bituminosa (control treatment), (ii) sowing of P. miliaceum for co-cultivation of both species, (iii) sowing of P. miliaceum and co-cultivation of both species in soil with compost, (iv) harvesting and elimination of the aerial part of the plants before sowing of P. miliaceum and (v) harvesting and incorporation to the soil of the aerial part of B. bituminosa before sowing of P. miliaceum.

Results and discussion

The results showed that the co-culture of both species favoured the growth of P. miliaceum, whilst incorporating the aerial part of the legume to the soil increased nitrogen concentration in P. miliaceum but reduced its growth. The use of compost improved both the growth and N uptake of P. miliaceum and did not inhibit nodulation in B. bituminosa. TE extractability in the soils and accumulation in the plants were rather low and very little affected by the addition of the amendments or by co-culture of species.

Conclusions

Nitrogen availability plays an important role in P. miliaceum growth in TE-contaminated mine soils. The addition of compost together with legume cultivation is proposed as an effective combination for the cultivation of P. miliaceum in these soils, as both plant growth and soil conditions were improved following this procedure.
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18.

Purpose

This study aimed at evaluating the acute effects of arsenic and zinc to the warmwater aquatic oligochaete Branchiura sowerbyi. Relative sensitivity with the coldwater species Tubifex tubifex was compared. Implications for the use of B. sowerbyi in the risk assessment of sediments in the tropics are discussed.

Materials and methods

Water-only (96 h) and sediment (14 days) toxicity tests were conducted with both species evaluating a concentration series of arsenic and zinc. The tests were conducted considering the environmental conditions in the natural habitat of T. tubifex (predominantly temperate) and B. sowerbyi (predominantly tropical). Both lethal and sublethal endpoints (autotomy of the posterior body parts, abnormal behavior and appearance) were determined in the tests. The lethal (LC10 and LC50) and effect (EC10 and EC50) concentrations were also determined to assess metal sensitivity for both species.

Results and discussion

Both test species were more sensitive to Zn than As in water-only tests, which is in agreement with previous studies evaluating the toxicity of these metals to aquatic oligochaetes. Sublethal effects were generally noted at concentrations lower than those leading to mortality. The warmwater oligochaete B. sowerbyi was more sensitive to both metals tested than the coldwater species T. tubifex.

Conclusions

Study findings support the need for using indigenous tropical species in risk assessments in the tropics. In addition, sublethal effect parameters should be included in toxicity testing with aquatic oligochaetes.
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19.

Purpose

Paulownia, one of the fastest growing broad-leaved tree species in the world, is widely distributed in the warm temperate regions of China. However, there are few commercial-scale Paulownia plantations, and there is only limited information available about the most suitable soil quality for Paulownia fortunei growth in mid-subtropical, Hunan Province, China.

Materials and methods

To understand the effect of the growth of P. fortunei on soil conditions, 25 soil property parameters under Paulownia plantations were studied in Hunan Province, China. Seventy-two standard plots of eight different stand types were analyzed by three statistical approaches to assess soil quality (SQ) in the different P. fortunei plantations.

Results and discussion

The results revealed that a majority of the soil characteristics when intercropping with oilseed rape and the pure P. fortunei (plantation III) were better than intercropping with Camellia oleifera, orange trees, and Cunninghamia lanceolata (Lamb.). Available calcium, available magnesium, available potassium, available phosphorus, soil thickness, slope, soil organic matter, available sulfur, available copper, dehydrogenase, and available zinc were selected as the minimum data set (MDS). The SQ index (SQI) showed that three classes for soil quality among the eight P. fortunei plantations ranged from 0.48 to 0.88 and these were correlated with standing volume (p?<?0.05).

Conclusions

From the results, we concluded that selected MDS indicators can describe the soil fertility quality of P. fortunei plantations, and that the relationship between SQI and standing volume has a biological significance. P. fortunei plantations intercropped with Camellia oleifera, orange trees, and Cunninghamia lanceolata (Lamb.) caused a deterioration in SQ, but intercropping oilseed rape and pure P. fortunei plantations produced an improvement in SQ.
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20.

Purpose

Evaluate the efficiency of Populus alba clone Villafranca in the uptake and translocation of Zn from contaminated soils.

Materials and methods

The effects of 48 days of zinc treatment (Zn t ) on the growth and the photosynthetic activities of P. alba L. clone Villafranca were studied using ZnSO4 (375 ppm per unit of soil dry weight) added in sand and peat moss substrate at the beginning of the treatment (T 0) and again after 30 days (T 1) in order to reach a target Zn concentration of 375 ppm at T 0 and 750 ppm at T 1 per unit of soil dry weight.

Results and discussion

Zn uptake in the different organs was analyzed after 30 (T 1) and 48 days (T 2) from the beginning of treatment, showing the following order: root ? leaves ≥ woody cutting = stem. The leaf area increased by 12 % in comparison to control plants at the end of second treatment (48 days). Cutting radial growth showed a high synchronicity in the growth rate fluctuation among control and Zn t plants, but a higher increase in radial diameter of Zn t cutting was observed starting from day 38 (after 8 days of second Zn t ) reaching after 48 days 38 % higher than control plants.

Conclusions

Although our data of leaf Zn concentration were in the range usually reported as toxic for plants, Villafranca clone in Zn t substrate were unaffected in terms of net CO2 assimilation and stomatal conductance to water vapor.
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