Profiling of PLFA: Implications for nonlinear spatial gradient of PCP degradation in the vicinity of Lolium perenne L. roots

An investigation was conducted using phospholipid fatty acids (PLFAs) profiles to follow the spatial response of the microbial community at the millimeter scale with the purpose of illustrating the mechanism of nonlinear spatial dependence of PCP degradation on the distance from the root surface in the rhizosphere of Lolium perenne L. A laminar rhizobox was designed to allow the harvest of intact layers of root compartment, near-(1, 2, 3, 4, 5 mm) and far-(>5 mm) rhizosphere soil from root surfaces without the removal of the root material itself. Lolium perenne L. was grown in environmental chambers for 53 days with soil spiked with 8.7 and 18 mg kg⁻¹ PCP. PLFA profiles were found to be affected by the distance from the rhizosphere, indicating a distance-dependent selective enrichment of competent species that may be responsible for efficient PCP degradation. In particular, the five fatty acids 16:1ω5, 16:0, i17:0, a17:0 and 10Me18:0 emerged as microbiological biomarkers that may be used for assessing phytoremediation processes of PCP in soil. Their synergistic effects were shown to be most responsive to the nonlinear spatial patterns of PCP degradation in the vicinity of Lolium perenne L. roots. The results suggest that root exudates induced modifications of microbial communities in the PCP contaminated rhizosphere and spatially modified the dominant species within these communities, resulting in the nonlinear PCP degradation pattern.     

Facilitation of pentachlorophenol degradation in the rhizosphere of ryegrass (Lolium perenne L.)

The phytoremediation of xenobiotics depends upon plant-microbe interactions in the rhizosphere, but the extent and intensity of these effects are currently unknown. To investigate rhizosphere effects on the biodegradation of xenobiotics, a glasshouse experiment was conducted using a specially designed rhizobox where ryegrass seedlings were grown for 53 days in a soil spiked with pentachlorophenol (PCP) at concentrations of 8.7±0.5 and 18±0.5 mg kg⁻¹ soil. The soil in the rhizobox was divided into six separate compartments at various distances from the root surface. Changes in PCP concentrations with increasing distance from the root compartment of the rhizobox were then assessed. The largest and most rapid loss of PCP in planted soil was at 3 mm from the root zone where total PCP decreased to 0.20 and 0.65 mg kg⁻¹, respectively with the two PCP treatments. The degradation gradient followed the order: near-rhizosphere>root compartment>far-rhizosphere soil zones for both concentrations where ryegrass was grown. In contrast, there was no difference in PCP concentration with distance in the unplanted soil. The increases in both soil microbial biomass carbon and the activities of soil urease and phosphatase were accompanied by the enhanced degradation of PCP, which was higher in the near-rhizosphere than far-rhizosphere soil. The results suggest that the effect of root proximity is important in the degradation of xenobiotics such as PCP in soil.     

Enhanced bioremediation of soil containing 2,4-dinitrotoluene by a genetically modified Sinorhizobium meliloti

The symbiotic nitrogen-fixing soil bacterium, Sinorhizobium meliloti, is well known for its ability to interact with the leguminous plant Medicago sativa L. It has, however, not been reported that this species possesses the capability to degrade toxic nitroaromatic compounds, such as 2,4-dinitrotoluene (DNT) which is commonly associated with the degradation of the explosive trinitrotoluene (TNT). In this study, the pJS1 DNT-biodegradative plasmid was genetically transferred to S. meliloti strain USDA 1936, which was confirmed by plasmid profile analysis. Several standard analytical and chemical tests including high performance liquid chromatography (HPLC), nitrite (NO₂) release assays, rhizosphere population and plant greenhouse studies were conducted to test the ability of S. meliloti to degrade 2,4-DNT. The possible presence of 2,4-DNT remaining in the treated soil was tested, and no 2,4-DNT had been absorbed by the soil. The pJS1-carrying recombinant strain DHK1 produced ‘ARC’ alfalfa plants that were almost 2-fold higher in shoot dry weight than that produced by the parent strain on soil containing 0.14 mM 2,4-DNT. The transconjugant strain DHK1 reduced significantly one-third more 2,4-DNT in both 0.14 and 0.28 mM contaminated soil, and in 0.55 mM contaminated soil it degraded 94% of the 2,4-DNT present. In liquid cultures, however, only about 4% reduction in 2,4-DNT concentrations was obtained in 10 days. We interpret the results as clearly establishing that genetic modification was successfully used, for the first time, to improve the capability of the symbiotic nitrogen-fixing soil bacterium S. meliloti DHK1 to bioremediate in situ 2,4-DNT-contaminated soil in the presence of alfalfa 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.     

Reductive transformation of pentachlorophenol on the interface of subtropical soil colloids and water

Ten soil colloids were obtained from three kinds of Fe-rich (> 50 g kg^− 1) subtropical soil parent materials (Basalt, Sandshale, and Quaternary Period Red Earth) collected in nine sites in Guangdong of China. Effect of the Fe-rich soil colloids and adding Fe(II) and oxalic acid on reductive dechlorination transformation of pentachlorophenol (PCP) were studied on colloids interfaces of reaction suspension. Mineralogical properties and specific surface area of the soil colloids were characterized by X-ray powder diffraction and Brunauer-Emmett-Teller (BET) methods, respectively. A series of reductive experiments were designed to determine PCP transformation and chloride ion release, and to calculate rate constant (k values) of pseudo first-order kinetics. Our results showed that reductive transformation of PCP occurred with k values from 0.007 to 0.057 d^− 1, and relevant chloride was released in the suspension of the ten soil colloids. Soil colloid developed from Basalt presented higher transformation rates (0.040-0.057 d^− 1) than that from Sandshale (0.007-0.033 d^− 1) and Quaternary Period red earth (0.012 d^− 1). Two paddy soil colloids developed from Sandshale (0.032-0.033 d^− 1) were more active than other three Sandshale soil colloids (0.007-0.011 d^− 1). The k values were significantly and positively correlated to the BET surface area (P < 0.01, n = 10). Addition of oxalic acid (0.022-0.231 d^− 1) or Fe(II) (0.029-0.256 d^− 1) into suspension of soil colloids gave arise to increase by 1.2-9.4 times in the k values. The release of chloride ion was simultaneously elevated. The enhancement of oxalic acid or Fe(II) on reductive transformation of PCP was attributed to increase of surface-bound Fe(II), which possess high reductive reactivity. The k values adding 1.0 mM oxalic acid were significantly and positively correlated to BET surface area and soil pH (P < 0.01), while k values adding 1.0 mM Fe(II) were related to total Fe (P < 0.001). The results may give new insight to understand the contribution of PCP abiotic reductive transformation in subtropical and tropical soils, and also in permeable reactive barriers.     

Brassica genotypes differ in growth, phosphorus uptake and rhizosphere properties under P-limiting conditions

Compared to other crops, Brassicas are generally considered to grow well in soils with low P availability, however, little is known about genotypic differences within Brassicas in this respect. To assess the role of rhizosphere properties in growth and P uptake by Brassicas, three Brassica genotypes (mustard, Brassica juncea cv Chinese greens and canola, Brassica napus cvs Drum and Outback) were grown in an acidic soil with low P availability at two treatments of added P: 25 and 100 mg P kg⁻¹ as FePO₄ (P25 and P100). The plants were harvested at the 6-leaf stage, at flowering and at maturity. Shoot and root dry weight (dry weight) and root length increased with time and were lower in P25 than in P100. In P25, shoot dry weight was lowest in Outback and highest in Chinese greens. In the P100 treatment, Chinese greens had a higher shoot dry weight than the two canola cultivars. Chinese greens had a lower root dry weight and root length at flowering and maturity than the canola genotypes in both P treatments. Irrespective of P treatment, shoot P concentration was lower in Chinese greens than in the two canola genotypes. Specific P uptake (μg P m⁻¹ root length) decreased with time. In P25, Chinese greens had the lowest specific P uptake at the 6-leaf stage but it was higher than in the two canola genotypes at flowering and maturity. In P100, Outback had the lowest specific P uptake. Available P in the rhizosphere (resin P) decreased over time with the greatest decrease from the 6-leaf stage to flowering. In P25, resin P in the rhizosphere was greatest in Chinese greens at the 6-leaf stage and flowering and smallest in Outback at flowering. Microbial P and acid phosphatase activity changed little over time, were not affected by P treatment and there were only small differences between the genotypes. The rhizosphere microbial community composition [assessed by fatty acid methyl ester (FAME) analysis] of Outback and Chinese greens differed from that of the other two genotypes at the 6-leaf stage and flowering, respectively. At maturity, all three genotypes had distinct microbial communities. Plant traits such as production of high biomass at low shoot P concentrations as well as the capacity to maintain high P availability in the rhizosphere by P mobilisation can explain the observed differences in plant growth and P uptake among the Brassica genotypes.     

Temperature sensitivity of soil respiration in different ecosystems in China

Understanding the sensitivity of soil respiration to temperature change and its impacting factors is an important base for accurately evaluating the response of terrestrial carbon balance to future climatic change, and thus has received much recent attention. In this study, we synthesized 161 field measurement data from 52 published papers to quantify temperature sensitivity of soil respiration in different Chinese ecosystems and its relationship with climate factors, such as temperature and precipitation. The results show that the observed Q₁₀ value (the factor by which respiration rates increase for a 10 °C increase in temperature) is strongly dependent on the soil temperature measurement depth. Generally, Q₁₀ significantly increased with the depth (0 cm, 5 cm, and 10 cm) of soil temperature measuring point. Different ecosystem types also exhibit different Q₁₀ values. In response to soil temperature at the depth of 5 cm, alpine meadow and tundra has the largest Q₁₀ value with magnitude of 3.05 ± 1.06, while the Q₁₀ value of evergreen broadleaf forests is approximately half that amount (Q₁₀ = 1.81 ± 0.43). Spatial correlation analysis also shows that the Q₁₀ value of forest ecosystems is significantly and negatively correlated with mean annual temperature (R = −0.51, P < 0.001) and mean annual precipitation (R = −0.5, P < 0.001). This result not only implies that the temperature sensitivity of soil respiration will decline under continued global warming, but also suggests that such acclimation of soil respiration to warming should be taken into account in forecasting future terrestrial carbon cycle and its feedback to climate system.     

Volatile organic compounds in the roots and rhizosphere of Pinus spp.

Plant roots normally release a complex mixture of chemicals which have important effects in the rhizosphere. Among these different root-emitted compounds, volatile isoprenoids have received very little attention, yet they may play important and diverse roles in the rhizosphere, contributing to the regulation of microbial activity and nutrient availability. It is therefore important to estimate their abundance in the rhizosphere, but so far, there is no reliable sampling method that can be used to measure realistic rates of root emissions from plants growing in field conditions, or even in pots. Here, we measured root content of volatile isoprenoids (specifically monoterpenes) for Pinus pinea, and explored the feasibility of using a dynamic bag enclosure method to measure emissions from roots of intact pot-grown plants with different degrees of root cleaning. We also investigated a passive diffusion method for exploring monoterpenes in soil at incremental distances from mature Pinus sylvestris trees growing in field conditions. Total monoterpene content of P. pinea roots was 415±50 μg g⁻¹ fresh wt in an initial screening study, and between 688±103 and 1144±208 μg g⁻¹ dry wt in subsequent investigations. Emissions from shaken-clean roots of intact plants and roots of intact plants washed to remove remaining soil after shaken-clean experiments were 119±14 and 26±5 μg g⁻¹ dry wt h⁻¹, respectively. Emissions from intact roots in soil-balls were an order of magnitude lower than from shaken-clean roots, and probably reflected the amount of emitted compounds taken up by physical, chemical or biological processes in the soil matrix surrounding the roots. Although monoterpene content was not significantly different in droughted roots, emission rates from droughted roots were generally significantly lower than from well-watered roots. Finally, passive sampling of monoterpenes in the soil at different distances from mature P. sylvestris trees in field conditions showed significantly decreasing sampling rates with increasing distance from the trunk. We conclude that it is feasible to measure volatile isoprenoid emissions from roots but the method of root preparation affects magnitude of measured emissions and therefore must be decided according to the application. We also conclude that the rhizosphere of Pinus species is a strong and previously un-characterized source of volatile isoprenoid emissions and these are likely to impact significantly on rhizosphere function.     

Potential for biodegradation of anthropogenic organic compounds at low temperature in boreal soils

The effect of temperatures of −2.5 to +20 °C on the biodegradation of concentrations 0.2-50 μg cm⁻³ of pentachlorophenol (PCP), phenanthrene, pyrene and 2,4,5-trichlorophenol (TCP) was studied in soils sampled from an agricultural field and a relatively pristine forest in Helsinki, Finland. At the temperatures simulating seasonal variation of boreal soil temperatures [Heikinheimo, M., Fougstedt, B., 1992. Statistic of Soil Temperature in Finland. Meteorological Publications 22. Finnish Meteorological Institute, Helsinki, Finland], the response of mineralization of PCP, phenanthrene and 2,4,5-TCP was the most effective in the rhizosphere fraction of the forest humus soil at the substrate concentrations of ?5 μg cm⁻³. In the control incubation, performed at constant temperature of +20 °C, the mineralization yields of the model pollutants were highest in the agricultural soil with the highest applied substrate concentration (50 μg cm⁻³). The results suggest that the high level of pollutant mineralization at +20 °C resulted from the apparent adaptation of the soil microbial community to the high substrate concentration. No such adaptation occurred when the soils were incubated at temperatures simulating the actual boreal soil temperatures. The present results stress the role of adjusting the incubation conditions to environmentally relevant values, when assessing biodegradation of anthropogenic organic compound in boreal soils.     

Biocontrol potential of native Trichoderma isolates against root-knot nematodes in West African vegetable production systems

Seventeen isolates of the free-living soil fungus Trichoderma spp., collected from Meloidogyne spp. infested vegetable fields and infected roots in Benin, were screened for their rhizosphere competence and antagonistic potential against root-knot nematodes, Meloidogyne incognita, in greenhouse pot experiments on tomato. The five isolates expressing greatest reproductive ability and nematode suppression in pots were further assessed in a typical double-cropping system of tomato and carrot in the field in Benin. All seventeen isolates were re-isolated from both soil and roots at eight weeks after application, with no apparent crop growth penalty. In pots, a number of isolates provided significant nematode control compared with untreated controls. Field assessment demonstrated significant inhibition of nematode reproduction, suppression of root galling and an increase of tomato yield compared with the non-fungal control treatments. Trichoderma asperellum T-16 suppressed second stage juvenile (J₂) densities in roots by up to 80%; Trichoderma brevicompactum T-3 suppressed egg production by as much as 86%. Tomato yields were improved by over 30% following the application of these biocontrol agents, especially T. asperellum T-16. Although no significant effects were observed on carrot galling and yield, soil J₂ densities were suppressed in treated plots, by as much as 94% (T. asperellum T-12), compared with the non-fungal controls. This study provides the first information on the potential of West-African Trichoderma spp. isolates for use against root-knot nematodes in vegetable production systems. The results are highly encouraging, demonstrating their strong potential as an alternative and complementary crop protection component.     

Glucosinolate and isothiocyanate concentration in soil following incorporation of Brassica biofumigants

The concentration of glucosinolates (GSLs) and isothiocyanates (ITCs) was monitored in soil following the incorporation of pulverised high and low GSL varieties of rape (Brassica napus) and mustard (Brassica juncea) biofumigant crops. The concentration of both GSLs and ITCs in soil was highest immediately (30 min) after incorporation and they could be detected for up to 8 and 12 d, respectively. Irrigating with 18 mm of water over 3 h had no effect on either GSL or ITC concentrations. The amounts detected were generally related to the amount of GSL added in the incorporated plant tissue. Maximum total GSL concentration detected in the soil was 13.8 and 22.8 nmol g⁻¹ for rape and mustard, respectively, representing 7% and 13% of the original GSL present in the incorporated tissues. The non-ITC liberating GSLs (predominately indolyl GSLs) were found at lower concentrations than ITC-liberating GSLs, but tended to persist longer in the soil. Maximum total ITC concentration was 21.6 nmol g⁻¹ and 90.6 nmol g⁻¹ for rape and mustard, respectively. Calculated ITC release efficiency was 26% and 56% for high GSL rape and mustard, respectively at the time of the highest ITC concentration measured. These are the first reported measurements of GSLs in soil following biofumigant incorporation. They indicate that a significant proportion of plant GSL can persist un-hydrolysed in the soil for several days following Brassica incorporation. Further investigations of plant treatment and incorporation methods to maximise ITC release are warranted.     

Enhanced microbial degradation of pentachlorophenol from soil in the presence of earthworms: Evidence of functional bacteria using DNA-stable isotope probing

This study investigated the effect of two earthworm species (Amynthas robustus Perrier and Eisenia fetida Savigny) on the soil microbial degradation of pentachlorophenol (PCP). PCP-degrading microbes were identified using DNA-stable isotope probing (SIP). The results showed that adsorption and fixing to soil particles and organic fractions dominated the fate of PCP in soil without any amendments. The inoculation of both earthworm species significantly enhanced soil PCP disappearance and basal respiration. The DNA-SIP results revealed that Klebsiella, Cupriavidus, Aeromonas, and Burkholderia spp. were present at higher relative abundances in [¹³C]-labeled-PCP-amended soil microcosms than [¹²C]-PCP-amended soil in the presence of A. robustus, indicating that these bacterial species were responsible for PCP assimilation. Cupriavidus and Aeromonas sp. were also detected in the earthworm gut before inoculation, and their relative abundance was affected by earthworms. These results demonstrated that earthworms can introduce functional bacteria into soils and increase the population of PCP-degrading bacteria, thereby accelerating soil PCP degradation.     

雷公藤根际微生物特征研究

根际微生物的代谢作用, 直接促进或抑制根的营养吸收和生长, 也影响根际土壤中的物质转化, 雷公藤根系发达且多与其他树种混交栽培, 其根际微生物活性对雷公藤的生长和土壤肥力均有不可忽视的影响。以福建省泰宁县3 种不同栽培模式雷公藤林(野生雷公藤林、杉木雷公藤混交林、厚朴雷公藤混交林)为研究对象, 通过稀释平板法测定3 种不同雷公藤林分根际土壤和非根际土壤中细菌、真菌、放线菌的数量。结果表明: 根际土的微生物数量大于非根际土的微生物数量; 3 种林分, 无论是在根际土壤中, 还是在非根际土壤中, 均表现为细菌数量＞放线菌数量＞真菌数量; 根际微生物(R)比非根际微生物(S)更活跃, 3 种林分的3 大类微生物的R/S 数量比值均大于1; 3 种林分的微生物活性表现为厚朴雷公藤混交林＞杉木雷公藤混交林＞野生雷公藤林, 表明混交方式可促进雷公藤根际微生物活性。     

Regional and local factors affecting diversity, abundance and activity of free-living, N2-fixing bacteria in Australian agricultural soils

N₂-fixation by free-living (diazotrophic) microorganism is a key process affecting ecosystem functioning in soils. Understanding drivers affecting diazotrophic community assemblages and activities may lead to management practices to increase primary production and/or environmental sustainability. We used PCR-DGGE to determine the fundamental relationships between diazotrophic community structure and in a wide range of soils across southern Australia. In addition qPCR, RT-qPCR and N₂-fixation (acetylene reduction) were used to investigate factors influencing gene abundance, expression and processes in similar soils with different agricultural inputs. Across 22 soils, the structural composition of the nifH community was significantly influenced by site (ANOSIM R = 0.876; P = 0.001). The effects of management practices were evident, and often larger than between-soil differences, but were only present at some sites. Differences in nifH communities between sites correlated to particulate organic carbon (POC; measured by mid-infrared spectroscopy) content of the soils (BIO-ENV test; ρ = 0.502; P = 0.001), but not other factors including total soil C. In 3 soils from the Murrumbidgee irrigation region of NSW, intensification of the farming systems was associated with increasing N₂-fixation (P < 0.05), except where rice was cultivated. N₂-fixation correlated either with nifH abundance or gene expression in soils, but not both. Our data shows that soil C is closely linked to diazotrophic ecology. Principally, the amount of C entering the soil system is directly related to the abundance and N₂-fixation activity of free-living bacteria. However, we also show that C in the POC pool has associative links to the genetic diversity of the soil diazotroph community. Given the importance of diversity and abundance of functional organisms in supporting ecosystem processes, we suggest that soil C inputs should be considered for both qualitative and quantitative properties when considering impacts on diazotrophic bacterial ecology.     

Behaviour of plant‐derived extracellular phytase upon addition to soil

The behaviour of phytase after addition to three soil types with different sorption capacities was investigated. Phytase was collected from the roots of transgenic Arabidopsis thaliana that express a phytase gene from Aspergillus niger. Phytase activity in solution and on the solid phase of the soil was monitored over time. Phytase added to the solution phase of a soil suspension (1:20, w/v) was almost completely lost within 10 min in all soil types, while phytase in non-soil controls remained active in solution. Phytase activity lost from solution was recovered on the soil solid phase, suggesting rapid adsorption of the enzyme. Adsorption of phytase was less in soil taken from the rhizosphere of transgenic plants expressing phyA, indicating that the rhizosphere environment may help maintain phytase activity in solution. The activity of adsorbed phytase declined with time at a rate 2-4 times slower than that in the absence of soil. Adsorption of phytase in soils was highest at pH 4.5, which is below the reported isoelectric point (pI) of the Aspergillus phytase. As soil pH increased, adsorption decreased until, at pH 7.5, all phytase was in solution. Where phytase remained in solution, activity was maintained for at least 8 d. In contrast, the activity of adsorbed phytase was increasingly inhibited with time, particularly at low pH. By increasing the pH in soil suspensions, phytase that had remained active on the soil solid phase for 28 d was almost totally desorbed. Rapid immobilisation of phytase in soil may limit its capacity to interact with phytate, and this may compromise the ability of transgenic plants which exude phytase from their roots to acquire P from endogenous soil phytate.     

Response of soil respiration to precipitation during the dry season in two typical forest stands in the forest-grassland transition zone of the Loess Plateau

Forest ecosystems on the Loess Plateau are receiving increasing attention for their special importance in carbon fixation and conservation of soil and water in the region. Soil respiration was investigated in two typical forest stands of the forest-grassland transition zone in the region, an exotic black locust (Robinia pseudoacacia) plantation and an indigenous oak (Quercus liaotungensis) forest, in response to rain events (27.7 mm in May 2009 and 19 mm in May 2010) during the early summer dry season. In both ecosystems, precipitation significantly increased soil moisture, decreased soil temperature, and accelerated soil respiration. The peak values of soil respiration were 4.8 and 4.4 μmol CO₂ m⁻² s⁻¹ in the oak plot and the black locust plot, respectively. In the dry period after rainfall, the soil moisture and respiration rate gradually decreased and the soil temperature increased. Soil respiration rate in black locust stand was consistently less than that in oak stand, being consistent with the differences in C, N contents and fine root mass on the forest floor and in soil between the two stands. However, root respiration (R_r) per unit fine root mass and microbial respiration (R_m) per unit the amount of soil organic matter were higher in black locust stand than in oak stand. Respiration by root rhizosphere in black locust stand was the dominant component resulting in total respiration changes, whereas respiration by roots and soil microbes contributed equally in oak stand. Soil respiration in the black locust plantation showed higher sensitivity to precipitation than that in the oak forest.     

Increased degradation of phenanthrene in soil by Pseudomonas sp. GF3 in the presence of wheat

A phenanthrene-degrading bacterial strain Pseudomonas sp. GF3 was examined for plant-growth promoting effects and phenanthrene removal in soil artificially contaminated with low and high levels of phenanthrene (0, 100 and 200 mg kg⁻¹) in pot experiments. Low and high phenanthrene treatments significantly decreased the growth of wheat. Inoculation with bacterial strain Pseudomonas sp. GF3 was found to increase root and shoot growth of wheat. Strain GF3 was able to degrade phenanthrene effectively in the unplanted and planted soils. Over a period of 80 days the concentration of phenanthrene in soil in which wheat was grown was significantly lower than in unplanted soil (p<0.05). At the end of the 80-d experiments, 62.2% and 42.3% of phenanthrene had disappeared from planted soils without Pseudomonas sp. GF3 when the phenanthrene was added at 100 and 200 mg kg⁻¹ soil, respectively, but 84.8% and 70.2% of phenanthrene had disappeared from planted soils with the bacterial inoculation. The presence of vegetation significantly enhances the dissipation of phenanthrene in the soil. There was no significant difference in soil polyphenol oxidase activities among the applications of 0, 100 and 200 mg kg⁻¹ of phenanthrene. However, the enzyme activities in planted and unplanted soils inoculated with the strain Pseudomonas sp. GF3 were significantly higher than those of non-inoculation controls. The bacterial isolate was also able to colonize and develop in the rhizosphere soil of wheat after inoculation.     

Diversity and antagonistic potential of Pseudomonas spp. associated to the rhizosphere of maize grown in a subtropical organic farm

Pseudomonas spp. are one of the most important bacteria inhabiting the rhizosphere of diverse crop plants and have been frequently reported as biological control agents (BCAs). In this work, the diversity and antagonistic potential of Pseudomonas spp. in the rhizosphere of maize cultivars Nitroflint and Nitrodent grown at an organic farm in Brazil was studied by means of culture-dependent and -independent methods, respectively. Sampling of rhizosphere soil took place at three different stages of plant development: 20, 40 and 106 days after sowing. A PCR-DGGE strategy was used to generate specific Pseudomonas spp. fingerprints of 16S rRNA genes amplified from total community rhizosphere DNA. Shifts in the relative abundance of dominant populations (i.e. PCR-DGGE ribotypes) along plant development were detected. A few PCR-DGGE ribotypes were shown to display cultivar-dependent relative abundance. No significant differences in diversity measures of DGGE fingerprints were observed for different maize cultivars and sampling times. The characterisation and assessment of the antagonistic potential of a group of 142 fluorescent Pseudomonas isolated from the rhizosphere of both maize cultivars were carried out. Isolates were phenotypically and genotypically characterised and screened for in vitro antagonism towards three phytopathogenic fungi and the phytopathogenic bacterium Ralstonia solanacearum. Anti-fungal activity was displayed by 13 fluorescent isolates while 40 isolates were antagonistic towards R. solanacearum. High genotypic and phenotypic diversity was estimated for antagonistic fluorescent Pseudomonas spp. PCR-DGGE ribotypes displayed by antagonists matched dominant ribotypes of Pseudomonas DGGE fingerprints, suggesting that antagonists may belong to major Pseudomonas populations in the maize rhizosphere. Antagonists differing in their genotypic and phenotypic characteristics shared the same DGGE electrophoretic mobility, indicating that an enormous genotypic and functional diversity might be hidden behind one single DGGE band. Cloning and sequencing was performed for a DGGE double-band which had no corresponding PCR-DGGE ribotypes among the antagonists. Sequences derived from this band were affiliated to Pseudomonas stutzeri and P. alcaligenes 16S rRNA gene sequences. As used in this study, the combination of culture-dependent and -independent methods has proven to be a powerful tool to relate functional and structural diversity of Pseudomonas spp. in the rhizosphere.     

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.     

Prey-predator dynamics in communities of culturable soil bacteria and protozoa: differential effects of mercury

We investigated whether the prey-predator dynamics of bacteria and protozoa were affected by inorganic mercury at concentrations of 0, 3.5 and 15 mg Hg(II) kg soil⁻¹. The amount of bioavailable Hg was estimated using a biosensor-assay based on the mer-lux gene fusion. The numbers of bacterial CFUs on the general medium 1/100 tryptic soy agar (TSA) were significantly decreased when the soil had been amended with Hg. In contrast, no effect was seen on the number of CFUs on the Pseudomonas-specific medium Gould's S1 agar. Protozoan numbers estimated by the most probable number (MPN) method with 1/100 TSB as growth medium were also negatively affected by Hg. The different fractions of protozoa were affected to different degrees suggesting that amoebae were less sensitive than slow-growing flagellates, which again were less sensitive than the fast-growing flagellates. In contrast, Hg did not induce any detectable changes in the diversity of flagellate morphotypes. In the treatment with 15 mg Hg kg⁻¹ a transiently increased number of bacteria was seen at day 6 probably concomitant with a decrease in the numbers of protozoa. This might indicate that Hg affected the prey-predator dynamics in communities of culturable bacteria and protozoa in soil. Furthermore, we showed that the number of Pseudomonas spp. was not affected by Hg whereas the number of bacteria growing on a general medium was.