The application of an organic amendment modifies the arbuscular mycorrhizal fungal communities colonizing native seedlings grown in a heavy-metal-polluted soil

A mesocosm experiment was conducted to investigate whether communities of arbuscular mycorrhizal (AM) fungi associated with roots of native (Piptatherum miliaceum, Retama sphaerocarpa, Psoralea bituminosa, Coronilla juncea, and Anthyllis cytisoides) and for comparison (Lolium perenne) seedlings in a heavy-metal-contaminated, semiarid soil were affected by the application of composted sugar beet waste. We also investigated whether there were relation between AMF diversity and metal concentration (Al, Cd, Cu, Fe, Mn, Pb and Zn) and total P in shoot as well as some soil parameters (total organic carbon and total N) when the SB waste was added to the soil. We analyzed a portion of approximately 795 base pairs of the small-subunit (SSU) rRNA gene by nested PCR, cloning, sequencing, and phylogenetic analyses. Twelve different AMF sequence types were distinguished: seven of these belonged to Glomus group A, one to Glomus group B, one to Diversispora, one to Archaeospora, and two to Paraglomus. The AM fungal populations colonizing roots in a heavy-metal-polluted soil were quite dependent on the host plant, the highest diversity values being obtained in authochtonous plants recognized as metallophytes, such as P. bituminosa, and in an allochtonous, invasive species (L. perenne). No significant correlation was found between AMF diversity and plant metal concentration and soil parameters. Excepting P. bituminosa, when sugar beet waste was added to soil, the populations of AM fungi in roots increased and the shoot metal concentrations decreased in all host plant species studied. Therefore, the addition of sugar beet waste can be considered a good strategy for the remediation and/or phytostabilization of mine tailing sites.     

Leachability of cadmium, lead, and zinc in a long-term spontaneously revegetated slag heap: implications for phytostabilization

Purpose

For abandoned slag heaps, the spontaneous establishment of a vegetation cover is usually considered beneficial as it represents a means of phytostabilization. However, for slag containing heavy metals, such a vegetation cover has a potential long-term effect on the fate of the metals. The objective of this study was to investigate how the long-term spontaneous revegetation of a slag heap can affect the fractionation and the leachability of Cd, Zn, and Pb.

Materials and methods

Soils from two plots covered by either Armeria maritima or Agrostis tenuis and a bare plot soil were sampled from a slag heap from a zinc smelting plant and characterized. The Community Bureau of Reference (BCR) sequential extraction scheme was adopted to determine the metal pools. The leachability of Cd, Pb, and Zn was assessed by means of a leaching column experiment.

Results and discussion

Long-term presence of a plant cover increased the proportion of Zn in the most mobile fraction and Pb in the fraction bound to organic matter. Cd distribution was relatively unaffected. Overall, the metal leachability was enhanced in the revegetated soils, notably due to higher organic anion release. However, responses of metal behavior to revegetation depended on the established plant species. The highest leachability of Cd was found in the soil covered by Agrostis tenuis, while the highest leachability of both Zn and Pb was observed in the soil below Armeria maritima.

Conclusions

Any remediation strategy for metal-rich waste dumps by phytostabilization should take into careful consideration the potential long-term mobilization effect of plant establishment on heavy metals. We conclude that, when using pioneer plants for phytostabilization purposes, preference should be given to pseudo-metallophyte over hyperaccumulator species.     

Metal Uptake by Spontaneous Vegetation in Acidic Mine Tailings from a Semiarid Area in South Spain: Implications for Revegetation and Land Management

Tailings are frequently a source of pollution in mining areas due to the spread of metals from their bare surfaces via wind or runoff water. Phytostabilization is an interesting and low-cost option to decrease environmental risks in these sites. In this study, an acidic mine tailing (pH 3?C4) located in a semiarid area in Southeast Spain and the spontaneous vegetation which grow on were investigated. Soil samples were taken to characterize metal contamination, and three plant species, Lygeum spartum, Piptatherum miliaceum, and Helichrysum decumbens, were sampled in order to determine plant uptake of metals. The rhizosphere pH of H. decumbens was measured to be 6.7, which was significantly higher than the bulk soil (pH 3). The electrical conductivity values were around 2?C5 dS m^?1. Total metal concentrations in soil were high (9,800 mg kg^?1 for Pb and 7,200 mg kg^?1 for Zn). DTPA-extractable Zn and Pb were 16% and 19% of the total amount, respectively. The three selected plant species accumulated around 2?C5 mg kg^?1 Cu in both shoots and roots. Zn concentration was 100 mg kg^?1 in P. miliaceum roots. DTPA-extractable Zn was positively correlated with Zn plant uptake. These plant species demonstrated to grow well in acid tailings taking up only low concentrations of metals and therefore are good candidates to perform further phytostabilization works.     

Stabilization of tannery sludge amended soil using Ricinus communis,Brassica juncea and Nerium oleander

Purpose

Characterization of tannery sludge (TS) for its plausible use in amelioration and phytoremediation of heavy metal rich TS treated soil by growing economically important plants (Ricinus communis, Brassica juncea and Nerium oleander).

Materials and methods

Treatments were prepared by amalgamation of TS (0, 5, 10, 20, 30, 50, 75, 100 %) with garden soil (GS). All treatments were analysed for chemical properties, total and DTPA (Diethylene triamine pentaacetic acid) extractable heavy metals (Cr, Pb, Cu, and Mn). Seed germination experiment was conducted; unvarying saplings were selected and planted in concerned pots and allowed to grow for 90 days in green house. At harvest, plant samples were washed with distilled water and used for determination of growth parameters (biomass of root, shoot and total biomass on dry weight basis) and metal accumulation in different parts of the plant. Translocation factor (TF) and bioconcentration factor (BCF) have been calculated to check the phytostabilization capability of studied plants.

Results and discussion

Application of TS in fixed quantity as an amendment resulted in significant improvements of GS characteristics (alkaline pH with high electrical conductivity, organic carbon, available NPK and heavy metals) and in treatments. DTPA extractable heavy metal concentrations were found very low and total heavy metal concentrations were also found under allowable range in control and treated soil (T-I to T-VI). The maximum seed germination percentage, plant growth, biomass production for all plant species were observed in T-III treatment (20 % TS + 80 % GS) with majority of the metal accumulation in underground part (BCF >1) and meagre translocation in aerial part (TF <1). From T-IV to T-VII treatment, accumulation of heavy metals in plant parts has generally increased; however, biomass has been tremendously decreased.

Conclusions

TS was found rich in NPK content with significant concentration of heavy metals. Pot growth experiment suggested amelioration of GS with specific quantity of 20 % TS can tremendously enhance the plant growth, help in the utilization of TS and can act as a substitute of synthetic fertilizer. Majority of the metals was accumulated in root part (BCF >1) and meagre translocation (TF <1) in aerial part, concludes R. communis and B. juncea could be suitable plant species to be grown in heavy metal rich TS treated soil, vis-à-vis for phytostabilization of heavy metals. In addition, these oil yielding and medicinal plants can also be used for phytoremediation of moderately contaminated tannery soils.

     

Response of <Emphasis Type="Italic">Piptatherum miliaceum</Emphasis> to co-culture with a legume species for the phytostabilisation of trace elements contaminated soils

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.

     

Survival of inocula and native AM fungi species associated with shrubs in a degraded Mediterranean ecosystem

Reconstitution of the potential of soil mycorrhizal inoculum is a key step in revegetation programs for semiarid environments. We tested the effectiveness of inoculation with native arbuscular mycorrhizal (AM) fungi or with an allochthonous AM fungus, Glomus claroideum, with respect to the growth of four shrub species, the release of mycorrhizal propagules in soil, within and outside the canopy, and the improvement of soil structural stability. Two years after outplanting, the mixture of native endophytes was more effective than, for Olea europaea subsp. sylvestris, Retama sphaerocarpa and Rhamnus lycioides, or equally as effective as, for Pistacia lentiscus, the non-native AM fungus Glomus claroideum, with respect to increasing shoot biomass and foliar NPK contents. The increases in glomalin concentration and structural stability produced by inoculation treatments in the rhizosphere soil of the all shrub species, except R. lycioides, ranged from about 55 to 173% and 13 to 21%, respectively. The mixture of native AM fungi produced the highest levels of mycorrhizal propagules in soil from the center of the canopy of P. lentiscus and R. lycioides, while plants of O. europaea and R. sphaerocarpa inoculated with G. claroideum had more mycorrhizal propagules than did those inoculated with the mixture of native fungi. The number of mycorrhizal propagules in soil outside the canopy of the four shrub species was 5-35 times higher in inoculation treatments than in soil of the non-inoculated plants.     

The significant contribution of mycorrhizal fungi and earthworms to maize protection and phytoremediation in Cd-polluted soils

Mycorrhizal fungi and earthworms can individually or interactively influence plant growth and heavy metal uptake. The influence of earthworms and arbuscular mycorrhizal (AM) fungi either alone or in combination on maize (Zea mays L.) growth and cadmium (Cd) uptake was investigated in a calcareous soil artificially spiked with Cd. Soils were contaminated with Cd (10 and 20 mg Cd kg⁻¹), inoculated or un-inoculated with the epigeic earthworm Lumbricus rubellus and two AM fungal species (Rhizophagus irregularis and Funneliformis mosseae) for two months of growth under greenhouse conditions. Generally, earthworms alone increased both shoot P uptake and biomass but decreased shoot Cd concentration and root Cd uptake. AM fungi individually often increased total maize P uptake, declined shoot Cd concentration, and consequently produced higher total biomass. However, R. irregularis enhanced shoot Cd uptake at low Cd level and root Cd uptake at high Cd level. In plants inoculated with F. mosseae species, earthworms increased shoot biomass and Cd uptake, decreased root biomass and Cd uptake at all Cd levels, and increased shoot Cd concentration at low Cd level. In plants colonized by R. irregularis species, however, earthworm addition decreased maize biomass only at high Cd level and root Cd concentration and total maize Cd uptake at both Cd levels. Earthworm activity decreased Cd transfer from the soil to maize roots at low Cd level, but this was counterbalanced in the presence of F. mosseae. Mycorrhizal symbiosis significantly reduced the transfer of Cd from roots to shoots, independence of earthworm effect. Overall, it is concluded that L. rubellus and AM fungi, in particular F. mosseae isolate, improved maize tolerance to Cd toxicity both individually and interactively by increasing plant growth and P nutrition, and restricting Cd transfer to the aboveground biomass. Consequently, the single and interactive effects of the two soil organisms might potentially be important not only in protecting maize plants against Cd toxicity, but also in Cd phytostabilization in soils polluted by this highly toxic metal.     

Relative Value of Phosphate Compounds in Reducing the Bioavailability and Toxicity of Lead in Contaminated Soils

Lead forms stable compounds with phosphate and the immobilized Pb becomes less available to soil biota. In this study, we tested the bioavailabilty of Pb using earthworms (Eisenia fetida) and plants after immobilization of Pb by a soluble P compound and an insoluble rock phosphate compound in the presence of phosphate-solubilizing bacteria (Enterobacter sp.). Rock phosphate in the presence of phosphate-solubilizing bacteria and a soluble P compound enhanced Pb immobilization as measured by NH₄NO₃-extractable Pb concentration, thereby reduced its bioavailability as evaluated by earthworm Pb loading and sunflower (Helianthus annuus) Pb uptake under greenhouse conditions. However, soluble P treatment increased the concentration of Pb in soil solution thereby inhibited the root elongation of mustard (Brassica hirta) seedlings. Sunflower plants in the Pb-spiked soil without P amendments showed symptoms of necrosis and stunting because of Pb toxicity. Both soluble and insoluble P treatments significantly increased shoot and root weight and decreased Pb concentration in shoot by more than 50% compared to the control. However, high Pb concentration in soil solution was found in soluble P treatment, which can be attributed to dissolved organic carbon–Pb complex formation, thereby increasing Pb mobility. The inoculation of phosphate-solubilizing bacteria can facilitate phytostabilization of Pb-contaminated site.     

Establishment of shrub species in a degraded semiarid site after inoculation with native or allochthonous arbuscular mycorrhizal fungi

The re-establishment of native shrub species in the Mediterranean basin serves to restore the characteristic biodiversity and to prevent the processes of erosion and desertification in semiarid areas. A field experiment was carried out in an abandoned semiarid agricultural Mediterranean area to assess the effectiveness of mycorrhizal inoculation, with a mixture of native arbuscular mycorrhizal (AM) fungi or an allochthonous AM fungus (Glomus claroideum), on the establishment of Olea europaea subsp. sylvestris L., Pistacia lentiscus L., Retama sphaerocarpa (L.) Boissier and Rhamnus lycioides L. seedlings in this area. One year after planting, shoot biomass of inoculated O. europaea and P. lentiscus seedlings was greater, by about 630% and 300%, respectively, than that of non-inoculated plants. Shoot biomass of G. claroideum-colonised R. sphaerocarpa plants was significantly greater than that of seedlings inoculated with the mixed native AM fungi after 12 months. The increase of R. lycioides growth due to inoculation with native AM fungi was significantly greater than that of G. claroideum-colonised seedlings during the same growth period. Inoculation with a mix of native AM fungi was the most effective treatment for increasing shoot biomass and N, P and K contents in shoot tissues of R. lycioides seedlings. The mixture of native AM fungi was the most effective with respect to colonisation of the roots of O. europaea and R. lycioides, but the native AM fungi and G. claroideum achieved similar levels of colonisation in P. lentiscus and R. sphaerocarpa. The use of native mycorrhizal potential as a source of AM inoculum may be considered a preferential inoculation strategy to guarantee the successful re-establishment of native shrub species in a semiarid degraded soil.     

Antioxidant activities and metal acquisition in mycorrhizal plants growing in a heavy-metal multicontaminated soil amended with treated lignocellulosic agrowaste

The plant growth, nutrient acquisition, metal translocation and antioxidant activities [ascorbate peroxidase (APX), glutatione reductase (GR), superoxide dismutase (SOD) and catalase (CAT)] were measured in plants growing in a heavy-metal (HM) multicontaminated soil inoculated with selected autochthonous microorganisms [arbuscular mycorrhizal (AM) fungus and/or plant growth promoting bacteria (PGPB)] and/or amended with an Aspergillus niger-treated agrowaste. The treated agrowaste on its own increased root growth by 296% and shoot growth by 504% compared with non-treated control plants. Both chemical and biological treatments, particularly when combined, enhanced plant shoot and root development. The stimulation effect on plant biomass was concomitant with increased AM colonization, P and K assimilation, and reduced metal translocation from soil to plant shoot. The treated residue, particularly through interactions with AM inoculation, produced the expected bioremediation effect, leading to enhanced plant development and successful rehabilitation of contaminated soil. The enhancement of CAT, APX and GR activities caused by AM inoculation suggests that AM colonization helped plants to limit oxidative damage to biomolecules in response to metal stress. The response of the plant's antioxidant activities to the amendment appears to be related to enhanced plant biomass production. The application of amendments and/or microbial inoculations to enhance plant growth and reduce metal translocation in multicontaminated soil could be a promising strategy for remediating HM pollution.     

Plant type differently promote the arbuscular mycorrhizal fungi biodiversity in the rhizosphere after revegetation of a degraded, semiarid land

It is suggested that the diversity of arbuscular mycorrhizal fungi (AMF) and their association with distinct plants species are crucial in the early stages of revegetation procedures since the AMF roots colonisation plays an important role improving plant establishment and growth. We carried out a study where we analyse the AMF community composition in the roots of Ephedra fragilis, Rhamnus lycioides, Pistacia lentiscus and Retama sphaerocarpa fourteen months after revegetation in a Mediterranean semiarid degraded area of southeast Spain in order to verify whether different plant species can variably promote the diversity of AM fungi in their rhizospheres after planted. We analysed a portion of approximately 795 bases pairs of the small-subunit ribosomal DNA by means of nested PCR, cloning, sequencing and phylogenetic analyses. Eight fungal sequence types belonging to Glomus group A and B and to the genus Paraglomus were identified. The different plant species had different AM fungal community composition. Thus, R. lycioides harboured the highest number of four fungal sequence types while from E. fragilis only two types could be characterized that were specific for this plant species. P. lentiscus and R. sphaerocarpa harboured each one three sequence types and two of them were shared. All AMF sequence types were found in the natural soil. These results show that one effective way of restoring degraded lands is to increase the number of plant species used, which would increase the AMF diversity in the soil and thus the below-ground, positive interactions.     

Do interactions with soil organisms mediate grass responses to defoliation?

Defoliation-induced changes in grass growth and C allocation are known to affect soil organisms, but how much these effects in turn mediate grass responses to defoliation is not fully understood. Here, we present results from a microcosm study that assessed the role of arbuscular mycorrhizal (AM) fungi and soil decomposers in the response of a common forage grass, Phleum pratense L., to defoliation at two nutrient availabilities (added inorganic nutrients or no added nutrients). We measured the growth and C and N allocations of P. pratense plants as well as the abundance of soil organisms in the plant rhizosphere 5 and 19 d after defoliation. To examine whether defoliation affected the availability of organic N to plants, we added ¹⁵N-labelled root litter to the soil and tracked the movement of mineralized ¹⁵N from the litter to the plant shoots.When inorganic nutrients were not added, defoliation reduced P. pratense growth and root C allocation, but increased the shoot N concentration, shoot N yield (amount of N in clipped plus harvested shoot mass) and relative shoot N allocation. Defoliation also reduced N uptake from the litter but did not affect total plant N uptake. Among soil organisms, defoliation reduced the root colonization rates of AM fungi but did not affect soil microbial respiration or the abundance of microbe-grazing nematodes. These results indicate that interactions with soil organisms were not responsible for the increased shoot N concentration and shoot N yield of defoliated P. pratense plants. Instead, these effects apparently reflect a higher efficiency in N uptake per unit plant mass and increased relative allocation of N to shoots in defoliated plants. The role of soil organisms did not change when additional nutrients were available at the moment of defoliation, but the effects of defoliation on shoot N concentration and yield became negative, apparently due to the reduced ability of defoliated plants to compete for the pulse of inorganic nutrients added at the moment of defoliation.Our results show that the typical grass responses to defoliation—increased shoot N concentration and shoot N yield—are not necessarily mediated by soil organisms. We also found that these responses followed defoliation even when the ability of plants to utilize N from organic sources, such as plant litter, was diminished, because defoliated plants showed higher N-uptake efficiency per unit plant mass and allocated relatively more N to shoots than non-defoliated plants.     

Nickel-tolerant Brevibacillus brevis and arbuscular mycorrhizal fungus can reduce metal acquisition and nickel toxicity effects in plant growing in nickel supplemented soil

The growth of clover (Trifolium repens ) and its uptake of N, P and Ni were studied following inoculation of soil with Rhizobium trifolii, and combinations of two Ni-adapted indigenous bacterial isolates (one of them was Brevibacillus brevis) and an arbuscular mycorrhizal (AM) fungus (Glomus mosseae). Plant growth was measured in a pot experiment containing soil spiked with 30 (Ni I), 90 (Ni II) or 270 (Ni III) mg kg⁻¹ Ni-sulphate (corresponding to 11.7, 27.6 and 65.8 mg kg⁻¹ available Ni on a dry soil basis). Single inoculation with the most Ni-tolerant bacterial isolate (Brevibacillus brevis) was particularly effective in increasing shoot and root biomass at the three levels of Ni contamination in comparison with the other indigenous bacterial inoculated or control plants. Single colonisation of G. mosseae enhanced by 3 fold (Ni I), by 2.4 fold (Ni II) and by 2.2 fold (Ni III) T. repens dry weight and P-content of the shoots increased by 9.8 fold (Ni I), by 9.9 fold (Ni II) and by 5.1 fold (Ni III) concomitantly with a reduction in Ni concentration in the shoot compared with non-treated plants. Coinoculation of G. mosseae and the Ni-tolerant bacterial strain (B. brevis) achieved the highest plant dry biomass (shoot and root) and N and P content and the lowest Ni shoot concentration. Dual inoculation with the most Ni-tolerant autochthonous microorganisms (B. brevis and G. mosseae) increased shoot and root plant biomass and subtantially reduced the specific absorption rate (defined as the amount of metal absorbed per unit of root biomass) for nickel in comparison with plants grown in soil inoculated only with G. mosseae. B. brevis increased nodule number that was highly depressed in Ni I added soil or supressed in Ni II and Ni III supplemented soil. These results suggest that selected bacterial inoculation improved the mycorrhizal benefit in nutrients uptake and in decreasing Ni toxicity. Inoculation of adapted beneficial microorganisms (as autochthonous B. brevis and G. mosseae) may be used as a tool to enhance plant performance in soil contaminated with Ni.     

Community structure of arbuscular mycorrhizal fungi associated with Robinia pseudoacacia in uncontaminated and heavy metal contaminated soils

The significance of arbuscular mycorrhizal fungi (AMF) in soil remediation has been widely recognized because of their ability to promote plant growth and increase phytoremediation efficiency in heavy metal (HM) polluted soils by improving plant nutrient absorption and by influencing the fate of the metals in the plant and soil. However, the symbiotic functions of AMF in remediation of polluted soils depend on plant–fungus–soil combinations and are greatly influenced by environmental conditions. To better understand the adaptation of plants and the related mycorrhizae to extreme environmental conditions, AMF colonization, spore density and community structure were analyzed in roots or rhizosphere soils of Robinia pseudoacacia. Mycorrhization was compared between uncontaminated soil and heavy metal contaminated soil from a lead–zinc mining region of northwest China. Samples were analyzed by restriction fragment length polymorphism (RFLP) screening with AMF-specific primers (NS31 and AM1), and sequencing of rRNA small subunit (SSU). The phylogenetic analysis revealed 28 AMF group types, including six AMF families: Glomeraceae, Claroideoglomeraceae, Diversisporaceae, Acaulosporaceae, Pacisporaceae, and Gigasporaceae. Of all AMF group types, six (21%) were detected based on spore samples alone, four (14%) based on root samples alone, and five (18%) based on samples from root, soil and spore. Glo9 (Rhizophagus intraradices), Glo17 (Funneliformis mosseae) and Acau3 (Acaulospora sp.) were the three most abundant AMF group types in the current study. Soil Pb and Zn concentrations, pH, organic matter content, and phosphorus levels all showed significant correlations with the AMF species compositions in root and soil samples. Overall, the uncontaminated sites had higher species diversity than sites with heavy metal contamination. The study highlights the effects of different soil chemical parameters on AMF colonization, spore density and community structure in contaminated and uncontaminated sites. The tolerant AMF species isolated and identified from this study have potential for application in phytoremediation of heavy metal contaminated areas.     

Organic-acid-producing,phytate-mineralizing rhizobacteria and their effect on growth of pigeon pea (Cajanus cajan)

Phytates represent a significant pool of organic phosphorus (Po) that is largely unavailable to plants. This study deals with phytate-mineralizing (PM), organic-acid-producing (OAP) rhizobacterial isolates, their characterization and their effect on plant growth. Their genetic diversity was assessed by 16S rRNA amplified ribosomal DNA restriction analysis (ARDRA) and selected isolates were identified by partial sequencing of 16S rRNA gene. Na-phytate and Po rich poultry farm manure (PFM) used as sources of phosphorus in semi-solid-agar (SSA) medium and soil respectively, for plant inoculation studies, where Cajanus cajan (pigeon pea) used as plant. Of thirty-nine rhizobacterial isolates, nineteen were proficient at releasing phosphate (Pi) (up to 85 μg/ml) from sparingly soluble calcium (Ca)-phytate and concomitantly decreasing the pH of minimal medium with 100 mM glucose from 8.0 to below 5. When the medium contained glycerol in place of glucose, Ca-phytate remained undissolved with no significant Pi released and no decline in pH. Genetic diversity of phytate-mineralizing (PM) rhizobacterial isolates suggests that the isolates mainly fall in two populations: acid-producing (AP) population (mainly represented by members of Enterobacteriaceae) and non-AP population. OAP-PM rhizobacterial isolates were identified as Citrobacter, Pantoea, Klebsiella and Enterobacter species. Organic acids (OAs) secreted by PM isolates were detected by HPLC, showed secretion of gluconic and acetic acids. Importance of OAs in Ca-phytate dephosphorylation was demonstrated in vitro using A. ficuum phytase. Gluconate and acetate additions enhanced phytase catalyzed dephosphorylation of Ca-phytate in vitro. Sonicated cell lysates of isolates showed significant Pi release from Ca-phytate compared to whole cells, indicating inaccessibility of Ca-phytate due to poor solubility. Selected isolates showed that they possess cell-associated acid phytase and modulators of phytase activity suggested that the enzymes are histidine acid phosphatase (HAP) type of phytase. OAP-PM isolates PP1 and DHRSS showed significant increase in dry shoot/root ratio and P content of shoot in Na-phytate containing semi-solid agar (SSA) medium, but isolate DHRSS did not increase dry shoot/root ratio in soil experiments containing poultry farm manure as source of P, although it significantly increased shoot P content of plants. The inoculation of isolates enhanced the shoot P content and dry shoot/root ratio, but did not increase the dry weight in SSA medium. It may be concluded that some OAP-PM rhizobacterial isolates that release P from Ca-phytate show increase in shoot P content in phytate containing SSA medium and in soils.     

Microbial community structure and function in a soil contaminated by heavy metals: effects of plant growth and different amendments

We studied the effects of in situ remediation of a heavy metal (HM) contaminated soil on some soil chemical properties, microbial function and microbial structural diversity after 18 months. The experiment was carried out at semifield scale in containers filled with HM contaminated soil from the Aznalcóllar mine accident (Southern Spain, 1998). The remediation measures consisted of the application of different amendments and/or establishment of a plant cover (Agrostis stolonifera L.). Seven treatments were established: four organic treatments (municipal waste compost (MWC), biosolid compost (BC), leonardite (LEO) and litter (LIT)), one inorganic treatment (sugar beet lime (SL)) and two controls (control with plant cover (CTRP) and control without plant cover (CTR)). Several soil chemical (pH, soluble HM, total organic C (TOC), water-soluble C (WSC) and available-P) and biochemical properties (microbial biomass C (MBC), MBC/TOC ratio and enzyme activities) were determined. Microbial community structure was studied by means of ARDRA (amplified ribosomal DNA restriction analysis). The SL, MWC and BC treatments were the most efficient to raise soil pH and decrease soluble HM concentrations. Total organic C was increased in the organic treatments by 2 to 4-fold, whereas water-soluble C was statistically similar in the CTRP, SL and the organic treatments, probably due to the presence of a root system in all these treatments. Available-P was also increased in the BC, SL and MWC treatments due to the higher P content of the amendments applied in these treatments. Soil microbial function was generally enhanced in the amended and CTRP treatments. The MWC, BC and SL treatments were particularly efficient to increase microbial biomass C, the MBC/TOC ratio and the dehydrogenase and aryl-sulphatase enzyme activities. These results could be attributed to the amelioration of some of the soil chemical properties: increase in soil pH and water-soluble C and decrease of HM soluble concentrations. ARDRA analyses showed changes in structural diversity in both the bacterial and fungal community under the different treatments. Fingerprinting patterns of the 16S rDNA obtained with Hinf-I and of the 18S rDNA with Hpa-II revealed higher similarity percentages among samples from the same treatment compared with samples from the other treatments. In addition, a higher similarity was found between samples from all treatments under the Agrostis influence. The use of certain amendments and/or a plant cover is important for in situ remediation of HM contaminated soils, since these practices can affect soil chemical properties, as well as the microbial community function and structure.     

Lead and Zinc Extraction Potential of Two Common Crop Plants, Helianthus Annuus and Brassica Napus

Phytoextraction is a remediation technology that uses plants to remove heavy metals from soil. The success of a phytoextraction process depends on adequate plant yield (aerial parts) and high metal concentrations in plant shoots. A pot experiment was conducted to investigate the combination effects of plants [sunflower (Helianthus annuus) and canola (Brassica napus)] with soil treatments (manure, sulfuric acid and DTPA). Treatments, including two plants and seven soil treatments, which applied according to completely randomized factorial design with three replications. The largest shoot dry weight biomass production occurred in manure treatments for both plants. The maximum shoot concentrations of Pb and Zn were 234.6 and 1364.4 mg kg^?1 respectively in three mmoles DTPA kg^?1 treatment of sunflower. Furthermore the results showed that sunflower had a higher extracting potential for removal of Pb and Zn from polluted soil.     

Suitability of four woody plant species for the phytostabilization of a zinc smelting slag site after 5 years of assisted revegetation

Purpose

Zinc smelting activity generates large volumes of highly toxic waste slags and poses a potential extreme environmental risk for the surrounding areas. The establishment of a vegetation cap for the phytostabilization of abandoned mine tailing heaps using plants is usually considered a beneficial approach. This study aimed to evaluate the suitability of phytostabilization of zinc smelting slag using four woody plants combined with organic amendments, to investigate the distribution of heavy metals in the slag–plant system, and to better understand how the direct revegetation of a zinc smelting slag site can influence the mobility and geochemical fraction of heavy metals.

Materials and methods

Slags were collected from the areas planted with vegetation (Arundo donax, Broussonetia papyrifera, Robinia pseudoacacia, and Cryptomeria fortunei) and a bare area in a zinc smelting waste slag site using an indigenous method. Physicochemical properties were determined with the usual procedures. The geochemical fraction and bioavailability of heavy metals was determined using the three-step modified European Community Bureau of Reference (BCR) sequential extraction and diethylene triamine pentaacetic acid (DTPA) sequential extraction schemes. Heavy metal concentrations (Cu, Pb, Zn, and Cd) in the slag and plant samples were also measured.

Results and discussion

Vegetation planted directly in the zinc smelting waste slag significantly enhanced the nutrient accumulation and reduced the bioavailability of heavy metals (Cu, Zn, and Cd) with the exception of A. donax for Zn and Cd. The presence of four woody plants increased the bioavailability of Pb. Sequential extraction revealed that revegetation reduced the acid-soluble extractable fraction and increased the fraction of heavy metals associated with the Fe/Mn oxy(hydr)oxides or organic matter. This is attributed to the establishment of plant-enhanced weathering of minerals in the waste slag that resulted in the formation of an amount of dissolved metals, and the amount of dissolved metals was partly redistributed into the soluble extractable fraction of the zinc smelting waste slag. The final concentration of metals (Cu, Pb, Zn, and Cd) in the soluble extractable fraction is dependent on the dynamics of metals induced by root activity in the rhizosphere. Much lower levels of heavy metals with lower translocation factors accumulated in the four woody plants than in the associated slags.

Conclusions

We conclude that the studied four woody plants showed a beneficial vegetation cover and phytostabilization potential within 5 years of revegetation. These woody plants have the potential for high heavy metal tolerance and low heavy metal accumulation. Therefore, these woody plants could be used for revegetation and phytostabilization of zinc smelting slag sites under field conditions.

     

Above- and below-ground plant inputs both fuel soil food webs

Soil food webs depend almost exclusively on plant derived resources; however, it is still subject to debate how plants affect soil biota. We tested the effects on soil decomposers of three components of soil inputs of plant species identity: presence of live plants (representing rhizodeposits), identity of shoot litter input and identity of root litter input; using all combinations of these for Trifolium pratense and Plantago lanceolata. We assessed impacts on soil microorganisms, Collembola, Oribatida and earthworms in a full-factorial greenhouse experiment. Species identity of shoot litter input had greatest effect on decomposers, following by species identity of live plant. Microbial carbon use efficiency and Oribatida density were significantly higher in the presence of T. pratense shoot litter input than in that of P. lanceolata shoot litter input, while earthworm body mass ratio was significantly higher in the presence of P. lanceolata plants than in that of T. pratense plants. Oribatida density was at minimum in the presence of P. lanceolata plants, shoot and root litter input, resulting in a significant three-way interaction and pointing to the relevance of all investigated plant input pathways. Live plant identity effects were not due to differences in living root biomass among species and treatments. Detrimental P. lanceolata effects may have been due to significantly lower N concentrations than in T. pratense tissue. Our results indicate that both above- and below-ground plant inputs into soil determine the performance of decomposers, and thus suggest due consideration of both types of inputs fueling soil food webs in future studies.     

Effect of corncob compost on plant growth in an acid red soil

Abstract

A comparison of corncob compost with lime on plant growth was studied in acid red soil with pH of 4.07. Lettuce, pea, and corn were selected as test plants for their varying tolerance to acid soil. The pot experiment compared six soil treatments and a check. Soil amendments were 1, 2, and 4 cmol calcium carbonate (CaCO₃) kg^‐1 and 5, 10, and 20 g corncob compost kg^‐1 soil. Results showed higher manganese (Mn) than aluminum (Al) content of the shoot in all check group plants. Reduced shoot Mn content increased shoot dry weight in all test plants, regardless of acid soil tolerance or soil treatment. The higher the test plant resistance to soil acidity, the weaker the detoxification effect of corncob compost was on Al uptake when compared with the check group. Liming was more effective at reducing shoot Mn content than corncob compost with the exception of the more acidity sensitive lettuce. Shoot phosphorus (P) content, however, increased with corncob compost from enhanced organic matter rates. Corncob compost treatments significantly increased shoot dry weight over liming in the acid soil. This study demonstrated an environmentally acceptable use for an agricultural waste.