Rhizosphere microbial community and Zn uptake by willow (Salix purpurea L.) depend on soil sulfur concentrations in metalliferous peat soils

On numerous occasions, rhizosphere microbial activities have been identified as a key factor in metal phytoavailability to various plant species and in phytoremediation of metal-contaminated sites. For soil bioremediation efforts in heavy metal contaminated areas, microbes adapted to higher concentrations of heavy metals are required. This study was a field survey undertaken to examine rhizosphere microbial communities and biogeochemistry of soils associated with Zn accumulation by indigenous willows (Salix purpurea L.) in the naturally metalliferous peat soils located near Elba, NY. Soil and willow leaf samples were collected from seven points, at intervals 18 m apart along a willow hedgerow, on four different dates during the growing season. Soil bacterial community composition was characterized by terminal restriction fragment length polymorphism (T-RFLP) analysis and a 16S clone library was created from the rhizosphere of willows and soils containing the highest concentrations of Zn. Bacterial community composition was correlated with soil sulfate, but not with soil pH. The clone library revealed comparable phylogenetic associations to those found in other heavy metal-contaminated soils, and was dominated by affiliations within the phyla Acidobacteria (32%), and Proteobacteria (37%), and the remaining clones were associated with a wide array of phyla including Actinobacteria, Gemmatimonadetes, Planctomycetes, Verrucomicrobia, Bacteriodetes, and Cyanobacteria. Diverse microbial populations were present in both rhizosphere and bulk soils of these naturally metalliferous peat soils with community composition highly correlated to the soil sulfate cycle throughout the growing season indicative of a sulfur-oxidizing rhizosphere microbial community. Results confirm the importance of soil characterization for informing bioremediation efforts in heavy metal contaminated areas and the reciprocity that microbial communities uniquely adapted to specific conditions and heavy metals may have on an ecosystem.     

The Influence of Heavy Metals and Mineral Nutrient Supply on Bituminaria bituminosa

With a view to the selection of plants for the re-vegetation of contaminated, semi-arid land, two populations of the perennial species Bituminaria bituminosa (Fabaceae) from the south of Spain were studied: one (“LA”) from a non-contaminated soil and the other (“C2”) from a similar soil having elevated total levels of Pb and Zn (1,112 and 4,249 μg g^?1, respectively). For sand-cultured plants receiving nutrient solution, flow cytometry showed that heavy metals, at the concentrations measured in aqueous extracts from contaminated soils, had only slight genotoxic effects on root tip cell nuclei. Both populations were also grown in both soils, in two pot experiments. In the first, shoot biomass of LA and C2 in the contaminated soil was decreased to similar extents, with respect to the “clean” soil. Tissue heavy metal concentrations were unlikely to have been phytotoxic, except in the case of shoot Zn for population LA, but there were tissue deficiencies of P and K for populations LA and C2, respectively. In the second pot assay, the stimulation of growth by NPK fertiliser confirmed that even though this soil had high total heavy metal levels, nutrient availability was the principal factor limiting growth. The lesser transport of heavy metals (Cd, Mn and Zn) to the shoot by the population from the contaminated site is a factor that should be considered when selecting B. bituminosa lines for the phytostabilisation of such sites.     

Uptake of Uranium by Lettuce (Lactuca sativa L.) in Natural Uranium Contaminated Soils in Order to Assess Chemical Risk for Consumers

Uranium mining activity in Cunha Baixa (Portugal) village has left a legacy of polluted soils and irrigation water. A controlled field experiment was conducted with lettuce (Lactuca sativa L.) in an agricultural area nearby the abandoned mine in order to evaluate uranium uptake and distribution in roots and leaves as well as ascertain levels of uranium intake by the local inhabitants from plant consuming. Two soils with different average uranium content (38 and 106 mg/kg) were irrigated with non-contaminated and uranium contaminated water (<20 and >100 μg/l). A non-contaminated soil irrigated with local tap water (<1 μg/l uranium) was also used as a control. Uranium in lettuce tissues was positively correlated with soil uranium content, but non-significant differences were obtained from contaminated soils irrigated with different water quality. Uranium in plants (dry weight) growing in contaminated soils ranged from 0.95 to 6 mg/kg in roots and 0.32 to 2.6 mg/kg in leaves. Lettuce bioconcentration is more related to available uranium species in water than to its uranium concentration. Translocated uranium to lettuce leaves corresponds to 30% of the uranium uptake whatever the soil or irrigation water quality. A maximum uranium daily intake of 0.06 to 0.12 μg/kg bodyweight day was estimated for an adult assuming 30 to 60 g/day of lettuce is consumed. Although this value accounts for only 10% to 20% of the recommended Tolerable Daily Intake for ingested uranium, it still provides an additional source of the element in the local inhabitants’ diet.     

Use of sawdust for soil ph amendments

Abstract

Changes in pH values during 12 weeks incubation in soils treated with acidified sawdust (ACD‐SD)‐treated soils ranged from 5.03 to 5.89, from 9.88 to 10.35 in soil treated with alkalized sawdust (ALK‐SD), and ranged from pH 6.88 to 7.35 in untreated sawdust‐amended soil. In unamended soil, pH values were 6.80 to 7.35. Bacterial populations over the 12 weeks in ACD‐SD‐treated soils increased from 5×10⁶ to 167×10⁶ colonies while bacterial populations in ALK‐SD‐treated soils increased from 2×10⁶ to 54×10⁶. Fungal populations increased from 6×10⁴ to 11,333×10⁴ colonies per gram soil in ACD‐SD treated soils over the 12 week incubation. Untreated sawdust and control soil did not result in any significant changes in the fungal populations.     

Remediation of Metal Contaminated Soil by Organic Metabolites from Fungi I—Production of Organic Acids

Investigations were made on living strains of fungi in a bioremediation process of three metal (lead) contaminated soils. Three saprotrophic fungi (Aspergillus niger, Penicillium bilaiae, and a Penicillium sp.) were exposed to poor and rich nutrient conditions (no carbon availability or 0.11 M d-glucose, respectively) and metal stress (25 µM lead or contaminated soils) for 5 days. Exudation of low molecular weight organic acids was investigated as a response to the metal and nutrient conditions. Main organic acids identified were oxalic acid (A. niger) and citric acid (P. bilaiae). Exudation rates of oxalate decreased in response to lead exposure, while exudation rates of citrate were less affected. Total production under poor nutrient conditions was low, except for A. niger, for which no significant difference was found between the poor and rich control. Maximum exudation rates were 20 µmol oxalic acid g^?1 biomass h^?1 (A. niger) and 20 µmol citric acid g^?1 biomass h^?1 (P. bilaiae), in the presence of the contaminated soil, but only 5 µmol organic acids g^?1 biomass h^?1, in total, for the Penicillium sp. There was a significant mobilization of metals from the soils in the carbon rich treatments and maximum release of Pb was 12% from the soils after 5 days. This was not sufficient to bring down the remaining concentration to the target level 300 mg kg^?1 from initial levels of 3,800, 1,600, and 370 mg kg^?1in the three soils. Target levels for Ni, Zn, and Cu, were 120, 500, and 200 mg kg^?1, respectively, and were prior to the bioremediation already below these concentrations (except for Cu Soil 1). However, maximum release of Ni, Zn, and Cu was 28%, 35%, and 90%, respectively. The release of metals was related to the production of chelating acids, but also to the pH-decrease. This illustrates the potential to use fungi exudates in bioremediation of contaminated soil. Nonetheless, the extent of the generation of organic acids is depending on several processes and mechanisms that need to be further investigated.     

Soil microorganisms antagonistic towards Rhizobium japonicum

Success in introducing Rhizobium japonicum strains into soil is related to their interaction with native microorganisms including some that are antagonistic. Actinomycetes, bacteria, fungi and rhizobiophages antagonistic towards strains of R. japonicum were counted directly using soil samples from field plots under different crop and soil management systems. The antagonistic actinomycete population varied from 1.3 × 10³ to 4.5 × 10⁵ g^?1 dry soil and ranged up to 90% of total actinomycetes. Soybean rhizosphere soil samples included antagonistic actinomycetes ranging up to 70% of total actinomycetes. The antagonistic bacterial population was less than 10% of total bacteria and the proportion did not vary significantly with crop or soil management practices. Antagonistic fungi were observed for many of the soils examined but they could not be counted. There were few rhizobiophages and they were found most frequently in soybean rhizospheres. Occasional bacterial and actinomycete colonies that stimulated growth of R. japonicum were randomly observed among the soil samples tested.     

Copper/Zinc Bioaccumulation and the Effect of Phytotoxicity on the Growth of Lettuce (<Emphasis Type="Italic">Lactuca sativa</Emphasis> L.) in Non-contaminated,Metal-Contaminated and Swine Manure-Enriched Soils

Copper/zinc bioaccumulation and the effect of phytotoxicity on the growth of lettuce (Lactuca sativa L.) were studied in plastic vessels containing (i) non-contaminated soil, (ii) copper-contaminated soils at concentrations of 75.0 and 125.0 mg kg^?1, (iii) zinc-contaminated soils at concentrations of 1200 and 2400 mg kg^?1, and (iv) soil enriched with swine manure. Copper and zinc concentrations in lettuce leaves were determined by flame atomic absorption spectrometry during 42 days of growth. Copper concentrations from 0.92 to 13.06 mg kg^?1 were found in lettuce leaves grown in copper-contaminated soils and zinc concentrations from 58.13 to 177.85 mg kg^?1 were found in lettuce leaves grown in zinc-contaminated soils. Copper and zinc concentrations in lettuce leaves grown in swine manure-enriched soils ranged from 0.82 to 8.33 and 0.68 to 13.27 mg kg^?1, respectively. Copper and zinc bioaccumulation caused a decrease in lettuce growth in metal-contaminated soils and an increase in phytotoxicity effects when compared to growth in non-contaminated and manure-enriched soils. These findings were confirmed by measuring leaf areas and biomasses. Copper was less toxic to lettuce than zinc due to the different concentrations in the soil. Lettuce growth and development was better in the swine manure-enriched soil than non-contaminated soil, which indicates that swine manure is a safe agricultural biofertilizer when used in appropriate amounts to avoid metal bioaccumulation in soil and plants.     

Biodegradation of hexachlorocyclohexane-isomers in contaminated soils

Several sites that are contaminated with isomers of the chlorinated insecticide hexachlorocyclohexane (HCH) are present across the globe and cause toxicity. For their bioremediation, we studied the degradation of HCH-isomers in contaminated soils by an isolate Pseudomonas aeruginosa ITRC-5. The degradation is optimal at 2 mg technical-HCH (t-HCH)/g soil, 15% water content, pH 8.0, temperature 28 °C and inoculum density 10⁶ colony forming unit/g soil. Under these conditions, from 5 kg soil, >98% α- and γ-HCH, 17% β-HCH and 76% δ-HCH are degraded after 15 days of incubation, which is accompanied with the release of 600 μg chloride/mg t-HCH. Concomitant to the degradation, a four-fold reduction in the toxicity of HCH-isomers to earthworm, Eisenia foetida, is also observed. Addition of ITRC-5 enhanced the degradation of soil-applied HCH-isomers in ‘open field’ conditions as well, and 97%, 43%, 94% and 77% of α-, β-, γ- and δ-HCH, respectively, are degraded after 12 weeks of incubation. Thus, the bacterium causes microbial degradation and detoxification of HCH-isomers, and can be used for the bioremediation of contaminated soils.     

Phenoxy herbicide degradation in soils: Quantitative studies of 2,4-D- and MCPA-degrading microbial populations

Microbial populations able to degrade 2,4-D (2,4-dichlorophenoxyacetate) and MCPA (4-chloro-2-methylphenoxyacetate) were enumerated by means of a most probable number (MPN) procedure in eight Natal soils not previously treated with these herbicides. Estimated 2,4-D-degrading populations ranged from 1.26 to 245.2 and MCPA-degrading populations from 0.34 to 1377 g^?1 dry soil; in seven of the soils the populations of these organisms were less than 40 and 30 g^?1, respectively. Such counts indicate that for the successful isolation of 2,4-D- or MCPA-degrading microorganisms from soil, at least 1 g dry weight of soil should be used for enrichment cultures. The 2,4-D-degrading organisms occurred among the aerobic soil bacteria detectable by plate count, at frequencies of only 1 in 30 × 10³ to 1 in 36 × 10⁶ and the MCPA-degrading organisms at frequencies of 1 in 5 × 10³ to 1 in 133 × 10⁶; the ease with which the herbicide-degrading organisms can be isolated from enriched soil cultures treated with 2,4-D or MCPA is evidence of their massive preferential proliferation in response to the herbicides.Log 2,4-D- and MCPA-degrading populations did not differ significantly in four soil samples, but in the others either the 2,4-D- or the MCPA-degrading population was dominant. The longer persistence of MCPA compared with that of 2,4-D could therefore not be ascribed to quantitative differences in the populations of MCPA- and 2,4-D-degrading soil microorganisms.No relationship was evident between the soil populations of 2,4-D- or MCPA-degrading microorganisms and aerobic soil bacteria, and variations of the three populations among the soil samples were not associated in any obvious way with the soil physical and chemical characteristics, except perhaps an association of the highest counts of herbicide-degrading organisms with a sugar cane soil of sandy texture and high C: N ratio.     

覆盖模式及小麦根系对土壤微生物区系的影响

采用平皿分离培养法研究了5种栽培模式和小麦根系对土壤细菌、真菌及放线菌数量的影响。连续2年的定位测定结果表明:覆膜有利于土壤微生物数量增加。5种栽培模式中,小麦根区、根外土壤细菌数量均以覆膜模式下最高,分别为116.8×106cfu·g-1和86.7×106cfu·g-1;土壤真菌和放线菌数量均以垄沟覆膜(垄上覆膜、垄沟播种)模式下最高,分别为3.0×103cfu·g-1、1.4×103cfu·g-1和18.9×105cfu·g-1、19.7×105cfu·g-1。不同模式下小麦根系对土壤细菌和真菌数量影响较大,表现为根区高于根外;而根系对放线菌影响较小,只有补灌和覆膜2种模式为根区高于根外。多重比较结果显示,覆膜与其他模式之间细菌数量差异极显著,根区土壤细菌和真菌数量与根外存在显著差异。覆盖和根系能大幅度增加根区细菌、真菌和放线菌的数量,强化小麦根区根外细菌和真菌的数量差异。     

Survival and persistence of genetically modified Sinorhizobium meliloti in soil

Studies were conducted to evaluate the survival and persistence of Sinorhizobium meliloti 104A14 and two acid phosphatase-negative mutants in Kirkland (fine, mixed, thermic Udertic Paleustolls) silt loam soils with various fertility levels, and to assess the impact of inoculation on nodule occupancy and soil microbial community structure in the inoculated alfalfa (Medicago sativa L.) rhizosphere. Recovery of the inoculated strains was 100% (in the order of 10⁸ cells g⁻¹ soil) immediately following inoculation to soils, but decreased from 10⁸ cells g⁻¹ soil to undetectable levels in a nutrient-poor soil within 32 days. In a nutrient-rich soil, approximately 2–3% (4.7–7.43×10⁶ cells g⁻¹ soil) of the mutants and 23% (5.84×10⁷ cells g⁻¹ soil) of the wild-type inocula persisted for more than 64 days. Survivability and persistence of the wild-type S. meliloti were significantly greater than that of the genetically modified acid phosphatase negative mutants in all the soils tested. The persistence and nodule occupancy of the introduced S. meliloti in sterile and non-sterile soils were also tested for two repeated alfalfa growth periods in the same plant growth units, with a 1 month interval in between and no additional inoculation for the second period. Nodule occupancy of the introduced S. meliloti in non-sterile soils ranged from 30 to 60% for the first period and 85 to 100% for the second period. Our results suggest that survival and persistence of S. meliloti was enhanced by alfalfa cultivation and increased soil fertility, but impaired by mutation of acid phosphatase genes regardless of phosphorus nutritional levels. Moreover, inoculation with genetically modified S. meliloti strain 104A14 promoted indigenous bacterial growth in soil (increased bacterial population from 1.4×10⁶ to 4.3×10⁶ cells g⁻¹ soil), but not the growth of fungi and yeast. However, inoculation of the wild-type S. meliloti or genetically modified mutants did not result in significant changes in microbial community structure as indicated by EP indices and ratios of r/K strategists.     

Survival of Pseudomonas solanacearum biovars 2 and 3 in soil: Effect of moisture and soil type

The survival of Pseudomonas solanacearum biovars 2 and 3 in three soils, a Nambour clay loam, a Beerwah sandy loam and a Redland Bay clay, was compared at pressure potentials of ?0.003, ?0.05 and ?0.15 kPa. The soils were inoculated with mutants of P. solanacearum biovars 2 and 3, resistant to 2000 μg streptomycin sulphate ml^?1 and their survival measured every 6 weeks for 86 weeks in the clay loam and clay and for 52 weeks in the sandy loam. Soil populations declined with the initial drying necessary to bring the soil moisture to the specific pressure potentials; the initial counts for biovar 2 varied between 0.20 and 2.00 × 10⁹ cfu g^?1 soil and for biovar 3 between 0.17 and 1.29 × 10⁹ cfu g^?1 soil.The population decline in soil maintained at a constant pressure potential was expressed as the rate of population decline. Biovar 2 declined more rapidly than biovar 3. The rate of population decline of each biovar at ?0.003 and ?0.05 kPa was greater in clay loam than in sandy loam and at all pressure potentials it was greater in clay loam and sandy loam than in clay. There was also a tendency for the rate of population decline of both biovars to decrease in the drier soil treatments.     

Changes in proteolytic bacteria in paddy soils in response to organic management

Proteolytic bacterial communities, which mineralize organic nitrogen, play a key role in agricultural systems. In this study, alkaline metalloprotease (apr) gene fragments from proteolytic bacteria were investigated in bulk and rhizosphere paddy soil from four fields under organic management (for 2, 3, 5, and 9 years), and from one field under conventional management (for 2 years). We analyzed the abundance and structure of the proteolytic bacterial communities using real-time quantitative PCR and denaturing gradient gel electrophoresis. Our results showed that the abundance of proteolytic bacteria ranged from 1.57?×?10⁸ to 8.02?×?10⁸?copies/g of soil. In addition, the abundance of the proteolytic bacteria in the paddy soils under organic management was significantly higher than those in the paddy soil under conventional management. Moreover, the gene copy numbers in the rhizosphere soils were significantly higher than those in the bulk soils. The abundance of proteolytic bacteria tended to increase with the duration of organic management, with the highest abundance being found in the soil that had been under organic management for 5 years. However, the proteolytic bacteria communities in the paddy soils were not significantly affected by management practices. Phylogeny analysis showed that all gel bands obtained represented genes from Pseudomonas. Additionally, correlation analysis and canonical correspondence analysis showed that C/N, C, and N were important factors that influenced the abundance and community structure of the proteolytic bacteria. These results suggest that proteolytic bacteria are indicators in organic management systems, depolymerize organic N and hence maintain soil sustainability.

Abbreviations: CM: conventional management; OM: organic management; DGGE: denaturing gradient gel electrophoresis; qPCR: real-time quantitative PCR detecting system; COFCC: China organic food certification center; CCA: canonical correspondence analysis     

The effect of concentrations and properties of phenanthrene, pyrene, and benzo(a)pyrene on desorption in contaminated soil aged for 1 year

Purpose

The choice and timing of microorganisms added to soils for bioremediation is affected by the dominant bioavailable contaminants in the soil. However, changes to the concentration of bioavailable PAHs in soil are not clear, especially when several PAHs coexist. This study investigated the effects of PAH concentration and chemical properties on desorption in meadow brown soil after a 1-year aging period, which could reflect changes of PAH bioavailability during bioremediation.

Materials and methods

Based on the percentage of different molecular weights in a field investigation, high-level contaminated soil (HCS) and low-level contaminated soil (LCS) were prepared by adding phenanthrene (PHE), pyrene (PYR) and benzo(a)pyrene (BaP) to uncontaminated meadow brown soil. The concentrations of HCS and LCS were 250 mg?kg^?1 (PHE, PYR, and BaP: 100, 100, and 50 mg?kg^?1) and 50 mg?kg^?1 (PHE, PYR, and BaP: 20, 20, and 10 mg?kg^?1) respectively. The soils were aged for 1 year, after which desorption was induced by means of a XAD-2 adsorption technique over a 96-h period.

Results and discussion

The range of the rapidly desorbing fraction (F _rap) for PHE, PYR, and BaP in HCS and LCS was from 1.9 to 27.8 %. In HCS, desorption of PYR was most difficult, and the rate constant of very slow desorption (K _vs) of PYR was 8 orders of magnitude lower than that of BaP, which had similar very slow desorbing fractions (49.8 and 50.5 %, respectively). However, in LCS, desorption of PYR was the easiest; the K_vs of PYR was 8–10 orders of magnitude higher than those of PHE and BaP. In HCS, the time scale for release of 50 % of the PAHs was ranked as BaP?>?PYR?>?PHE, while in LCS this was BaP?>?PHE?>?PYR.

Conclusions

The combined effect of PAH concentrations and properties should be taken into account during desorption. The desorption of PAH did not always decrease with increasing molecular weight, and the desorption of four-ring PAHs might be special. These results are useful for screening biodegrading microbes and determining when they should be added to soils based on the dominant contaminants present during different periods, thus improving the efficiency of soil bioremediation.     

Bioavailability,fractionation of pb and Zn in the rhizosphere of sunflower in chelators-amended contaminated soil

Assisted phytoremediation procedures have been widely employed as soil removal instrument of heavy metals from contaminated soils. Rhizosphere processes have a major impact on pb and Zn availability and its fractions in soils. The present study evaluates the effects of EDTA, citric acid (CA) and poultry manure extract (PME) on bioavailability and fractionation of pb, Zn in both the rhizosphere of sunflower (Helianthus annuus L.) and bulk soil. EDTA and CA were added to soils at the rates of 0, 0.5 and 1 mmol kg^?1 soil and PME at 0, 0.5 and 1 g kg^?1 soil as factorial in a completely randomized pattern with three replicates in greenhouse condition. Results showed that chelator application had a significant impact (p < 0.05) on pb, Zn extraction by different extractants and its fractions in soils. The order of concentrations of pb, Zn present in different fractions in soil treated by chelators was: oxides-bounded fraction > residual fraction > OM-bounded fraction > carbonate-bounded fraction > exchangeable fraction. Biochemical soil characteristics in the sunflower rhizosphere change resulting from its roots contributing to pb, Zn decline in mobile soil fractions, and change in soil pb, Zn fractions that are generally regarded as more stable.     

Resistance of microbial populations in DDT-contaminated and uncontaminated soils

One DDT-contaminated soil and two uncontaminated soils were used to enumerate DDT-resistant microbes (bacteria, actinomycetes and fungi) by using soil dilution agar plates in media either with 150 μg DDT ml⁻¹ or without DDT at different temperatures (25, 37 and 55°C). Microbial populations in this study were significantly (p<0.001) affected by DDT in the growth medium. However, the numbers of microbes in long-term contaminated and uncontaminated soils were similar, presumably indicating that DDT-resistant microbes had developed over a long time exposure. The tolerance of isolated soil microbes to DDT varied in the order fungi>actinomycetes>bacteria. Bacteria from contaminated soil were more resistant to DDT than bacteria from uncontaminated soils. Microbes isolated at different temperatures also demonstrated varying degrees of DDT resistance. For example, bacteria and actinomycetes isolated at all incubation temperatures were sensitive to DDT. Conversely fungi isolated at all temperatures were unaffected by DDT.     

Inoculation of seeds and soil with basidiospores of mycorrhizal fungi

It was demonstrated that basidiospores of the fungus Rhizopoyon luteolus, mycorrhizal for Pinus radiata, could be used successfully as seed inoculum after freeze-drying and storage for 3 months at 22°C, provided the inoculum level was increased 100-fold. Spore inoculum applied to seed could be held dry for at least 2 days before planting provided inoculum was increased 10-fold. On sowing freshly inoculated seed to sterile soil, 3 × 10³ basidiospores/seed were adequate for infection but maximum mycorrhizal infection occurred with 3 × 10⁴ spores/seed.A dose-response curve was obtained for mycorrhizal infection when basidiospores were applied to soil. As few as 100 spores/290 cm³ pot were sufficient for mycorrhizal infection although infection increased with greater spore dose to a maximum of 10⁵ spores/pot. Plant growth response was related to intensity of infection. It is suggested that the percentage germination of basidiospores in the rhizosphere may be considerably greater than those reported in studies with synthetic medium. A rhizosphere effect on germination of basidiospores was demonstrated and a method developed to facilitate studies of spore germination in the rhizosphere.     

Influence of the sunflower rhizosphere on the biodegradation of PAHs in soil

Reduced bioavailability to soil microorganisms is probably the most limiting factor in the bioremediation of polycyclic aromatic hydrocarbons PAH-polluted soils. We used sunflowers planted in pots containing soil to determine the influence of the rhizosphere on the ability of soil microbiota to reduce PAH levels. The concentration of total PAHs decreased by 93% in 90 days when the contaminated soil was cultivated with sunflowers, representing an improvement of 16% compared to contaminated soil without plants. This greater extent of PAH degradation was consistent with the positive effect of the rhizosphere in selectively stimulating the growth of PAH-degrading populations. Molecular analysis revealed that the increase in the number of degraders was accompanied by a dramatic shift in the structure of the bacterial soil community favoring groups with a well-known PAH-degrading capacity, such as Sphingomonas (α-Proteobacteria), Commamonas and Oxalobacteria (β-Proteobacteria), and Xhanthomonas (γ-Proteobacteria). Other groups that were promoted for which degrading activity has not been reported included Methylophyllus (β-Proteobacteria) and the recently described phyla Acidobacteria and Gemmatimonadetes. We also conducted mineralization experiments on creosote-polluted soil in the presence and absence of sunflower root exudates to advance our understanding of the ability of these exudates to serve as bio-stimulants in the degradation of PAHs. By conducting greenhouse and mineralization experiments, we separated the chemical impact of the root exudates from any root surface phenomena, as sorption of contaminants to the roots, indicating that sunflower root exudates have the potential to increase the degradation of xenobiotics due to its influence on the soil microorganisms, where sunflower root exudates act improving the availability of the contaminant to be degraded. We characterized the sunflower exudates in vitro to determine the total organic carbon (TOC) and its chemical composition. Our results indicate that the rhizosphere promotes the degradation of PAHs by increasing the biodegradation of the pollutants and the number and diversity of PAH degraders. We propose that the biostimulation exerted by the plants is based on the chemical composition of the exudates.     

The number of microorganisms and the microbiological activity of human-modified cryogenic pale soils of Yakutia

Specific features of the microbial population—the high number of all the groups of microorganisms (1.6 × 10³–3.5 × 10⁷) similar to their abundance in steppe soils of the Transbaikal region and the distribution of microorganisms along the soil profile (without a decrease in their number with depth)—were revealed in a cryogenic weakly solodized loamy sandy pale soil. Unlike the soils of the Transbaikal region and central Russia, where bacteria, as a rule, are accumulated in the litter and upper soil horizons, in the undisturbed and weakly disturbed soils of Yakutia, the number of microorganisms is rather high within the whole soil profile. In the strongly disturbed agropale soil of croplands, the distribution of microorganisms is the same as in the soils of the Transbaikal region and European Russia. In the cryogenic soils studied, actinomycetes predominated, and their number varied from 47 000 to 35 000 000 CFU/g of soil. The number of microorganisms positively (r = 1) correlated with the soil moisture.     

Release, movement and recovery of 3,4-dimethylpyrazole phosphate (DMPP), ammonium, and nitrate from stabilized nitrogen fertilizer granules in a silty clay soil under laboratory conditions

 The composition of soil microbiota in four heated (350  °C, 1 h) soils (one Ortic Podsol over sandstone and three Humic Cambisol over granite, schist or limestone) inoculated (1.5 μg chlorophyll a g^–1 soil or 3.0 μg chlorophyll a g^–1 soil) with cyanobacteria (Oscillatoria PCC9014, Nostoc PCC9025, Nostoc PCC9104, Scytonema CCC9801, and a mixture of the four) was studied by cultural methods. The aims of the work were to investigate the potential value of cyanobacteria as biofertilizers for accelerating soil recolonization after fire as well as promoting microbiotic crust formation and to determine the microbial composition of such a crust. The inoculated cyanobacteria proliferated by 5 logarithmic units in the heated soils which were colonized very quickly and, after 2 months of incubation, the cyanobacterial filaments and associated fungal hyphae made up a matrix in which surface soil particles were gathered into crusts of up to 1.0 cm in thickness. These crusts were composed, on average, of 2.5×10¹⁰ cyanobacteria, 2.8×10⁶ algae, 6.1×10¹⁰ heterotrophic bacteria (of which 1.2×10⁸ were acidophilic, 1.3×10⁶ were Bacillus spp. and 1.5×10⁸ were actinomycetes) and 77.8 m fungal mycelium (1.4×10⁶ were fungal propagules) g^–1 crust. Counts of most microbial groups were positively correlated to cyanobacterial numbers. The efficacy of treatment depended on both the class of inoculum and the type of soil. The best inoculum was the mixture of the four strains and, whatever the inoculum used, the soil over lime showed the most developed crust followed by the soils over schist, granite and sandstone; however, the latter was comparatively the most favoured by the amendment. In the medium term there were no significant differences between the two inocula rates used. Biofertilization increased counts of cyanobacteria by 8 logarithmic units while heterotrophic bacteria, actinomycetes, algae and fungal propagules rose by >4 logarithmic units, acidophilic bacteria and Bacillus spp. by around 3 logarithmic units and fungal mycelia showed an 80-fold increase. The results showed that inoculation of burned soils with particle-binding diazotrophic cyanobacteria may be a means of both improving crust formation and restoring microbial populations. Received: 8 March 2000