Using soil bacteria to facilitate phytoremediation

In the past twenty years or so, researchers have endeavored to utilize plants to facilitate the removal of both organic and inorganic contaminants from the environment, especially from soil. These phytoremediation approaches have come a long way in a short time. However, the majority of this work has been done under more controlled laboratory conditions and not in the field. As an adjunct to various phytoremediation strategies and as part of an effort to make this technology more efficacious, a number of scientists have begun to explore the possibility of using various soil bacteria together with plants. These bacteria include biodegradative bacteria, plant growth-promoting bacteria and bacteria that facilitate phytoremediation by other means. An overview of bacterially assisted phytoremediation is provided here for both organic and metallic contaminants, with the intent of providing some insight into how these bacteria aid phytoremediation so that future field studies might be facilitated.     

A mathematical model of phytoremediation for petroleum-contaminated soil: model development

We present a simple model for root length density that combines the generally accepted spatial (exponential decrease with depth) and temporal (sinusoidal) variability of root length. Parameters in this model for root length density can be determined from assumed or measured information regarding the annual biomass turnover, maximum standing biomass, and maximum depth of root penetration. The root length density model, coupled with information regarding the average root lifespan, gives specific root growth and senescence functions that are the forcing functions for the phytoremediation model. We present a screening level mathematical model for phytoremediation that accounts for the growth and senescence of roots in the system. This is an important factor for recalcitrant, immobile compounds found in weathered crude oil contaminated soils. The phytoremediation model is based on variable volume compartments that have individual first-order degradation rate constants; as the roots move through the soil, the soil cycles through the rhizosphere zone, decaying root zone and bulk soil zone. Thus, although the oil is immobile, as the roots penetrate through the soil the oil is brought into contact with the rhizosphere.     

Plant--rhizosphere-microflora association during phytoremediation of PAH-contaminated soil

The capability of plants to promote the microbial degradation of pollutants in rhizosphere soil is a principal mechanism of phytoremediation of PAH-contaminated soil. The formation of a specific rhizosphere microbocenosis with a high degradative potential toward contaminants is largely determined by plant species. The comparative PAH-degradation in unplanted soil and in soil planted with reed (Phragmites australis) and alfalfa (Medicago sativa) was studied in pot experiments during 2 years. Both alfalfa and reed successfully remediated contaminated soil by degrading 74.5 and 68.7% of PAHs, respectively. The study of the rhizosphere, rhizoplane, and unplanted-soil microflora in experimental pots showed that alfalfa stimulated the rhizosphere microflora of PAH-contaminated soil more effectively than did reed. Alfalfa clearly enhanced both the total number of microorganisms (1.3 times, according to fluorescence microscopy data) and the rate of the PAH-degrading population (almost seven times, according to plate counting). The degradative potential of its rhizosphere microflora toward PAHs was higher than the degradative activity of the reed rhizosphere. This study provides relevant information for the successful application of alfalfa to phytoremediate PAH-contaminated soil.     

Reclamation of tannery polluted soil through phytoremediation

The huge volume of sludge emanating from the tannery effluent treatment plants poses a serious environmental problem. Phytoremediation is an emerging technology in which the plants are employed to reclamate the contaminated soil strewn with heavy metals (metalloids) and toxic compounds. This work focuses the impact of application of tannery sludge on biochemical properties of 6 months old tree saplings of Azadirachta indica A. Juss. (Neem), Melia azedarach Linn. (Wild Neem) and Leucaena leucocephala (Lam) de Wit (Subabool) raised over the tannery sludge in an attempt to use these plants for phytoremediation. The plants raised over the garden soil served as the control. The porosity and water holding capacity of the tannery sludge were higher. The plant growth supporting elements such as Ca, total N₂, NO₃ and Mg were higher in the sludge. The plants raised over the sludge were found to be dark green with increased morphometric parameters. Electrophoretic profile revealed amplification of a few polypeptides (100, 105, 49 and 55 KDa). The levels of biomolecules and the CO₂ absorption increased in 6 months old plants. There was a significant uptake and transport of chromium in all the three tree species suggesting that these plants could be employed in phytoremediation of soils contaminated with heavy metals.Key words: Azadirachta indica (Neem), CO₂ uptake, Leucaena leucocephala (Subabool), Melia azedarach (Wild Neem), metalloids, Phytoremediation, Stress biomolecules, Tannery sludge     

Screening of plants for phytoremediation of oil-contaminated soil

Several species of ornamental flowering plants were evaluated regarding their phytoremediation ability for the cleanup of oil-contaminated soil in Japanese environmental conditions. Thirty-three species of plants were grown in oil-contaminated soil, and Mimosa, Zinnia, Gazania, and cypress vine were selected for further assessment on the basis of their favorable initial growth. No significant difference was observed in the above-ground and under-ground dry matter weight of Gazania 180 days after sowing between contaminated and non-contaminated plots. However, the other 3 species of plants died by the 180^th day, indicating that Gazania has an especially strong tolerance for oil-contaminated soil. The total petroleum hydrocarbon concentration of the soils in which the 4 species of plants were grown decreased by 45–49% by the 180^th day. Compared to an irrigated plot, the dehydrogenase activity of the contaminated soil also increased significantly, indicating a phytoremediation effect by the 4 tested plants. Mimosa, Zinnia, and cypress vine all died by the 180^th day after seeding, but the roots themselves became a source of nutrients for the soil microorganisms, which led to a phytoremediation effect by increase in the oil degradation activity. It has been indicated that Gazania is most appropriate for phytoremediation of oil-contaminated soil.     

A mathematical model of phytoremediation for petroleum contaminated soil: sensitivity analysis

Phytoremediation is an attractive treatment technology for many contaminated sites due to its cost effectiveness and public acceptance. We present a sensitivity analysis of important parameters from a screening level model for phytoremediation by grass species of weathered petroleum-contaminated sites. The conceptual framework is that root movement through contaminated soil will enhance contaminant biodegradation by providing a local environment more favorable for petroleum degrading microorganisms—the so-called rhizosphere effect. Common questions in phytoremediation are, “What species should be planted?” and “What management practices should be followed?” These choices may affect degradation kinetics, root biomass (and therefore rhizosphere volume), and the root turnover. Important model parameters are the rate constants, rhizosphere volume, and the rate of root turnover. We present a sensitivity analysis with the aim of identifying the most important factors for improving phytoremediation effectiveness. For simulations of the phytoremediation of weathered diesel range organics, our results indicate that annual species, with higher root turnover, are preferred over perennial species with the caveat of equal degradation rate constants, that is, no species-dependent effects. In addition, the results suggest that the management of nonrhizosphere soil could play an important role in the overall effectiveness of phytoremediation. Finally, the effect of increasing root biomass or increasing the rhizosphere thickness is approximately equivalent with respect to the ultimate removal of the contaminants.     

Understanding plant-microbe interactions for phytoremediation of petroleum-polluted soil

Plant-microbe interactions are considered to be important processes determining the efficiency of phytoremediation of petroleum pollution, however relatively little is known about how these interactions are influenced by petroleum pollution. In this experimental study using a microcosm approach, we examined how plant ecophysiological traits, soil nutrients and microbial activities were influenced by petroleum pollution in Phragmites australis, a phytoremediating species. Generally, petroleum pollution reduced plant performance, especially at early stages of plant growth. Petroleum had negative effects on the net accumulation of inorganic nitrogen from its organic forms (net nitrogen mineralization (NNM)) most likely by decreasing the inorganic nitrogen available to the plants in petroleum-polluted soils. However, abundant dissolved organic nitrogen (DON) was found in petroleum-polluted soil. In order to overcome initial deficiency of inorganic nitrogen, plants by dint of high colonization of arbuscular mycorrhizal fungi might absorb some DON for their growth in petroleum-polluted soils. In addition, through using a real-time polymerase chain reaction method, we quantified hydrocarbon-degrading bacterial traits based on their catabolic genes (i.e. alkB (alkane monooxygenase), nah (naphthalene dioxygenase) and tol (xylene monooxygenase) genes). This enumeration of target genes suggests that different hydrocarbon-degrading bacteria experienced different dynamic changes during phytoremediation and a greater abundance of alkB was detected during vegetative growth stages. Because phytoremediation of different components of petroleum is performed by different hydrocarbon-degrading bacteria, plants' ability of phytoremediating different components might therefore vary during the plant life cycle. Phytoremediation might be most effective during the vegetative growth stages as greater abundances of hydrocarbon-degrading bacteria containing alkB and tol genes were observed at these stages. The information provided by this study enhances our understanding of the effects of petroleum pollution on plant-microbe interactions and the roles of these interactions in the phytoremediation of petroleum-polluted soil.     

汞污染土壤植物修复中转基因技术的应用

汞是一种全球性污染物.随着工业的发展、汞矿开采的加速,土壤汞污染日益严峻,给生态环境和人类生活带来了严重的威胁.因此,汞污染土壤的修复问题受到了广大学者的关注.针对受污染的土壤修复问题,植物修复技术具有成本低和不会给环境带来新的危害等特点.本文对目前国内外汞污染土壤修复植物修复技术的研究进行了综述,介绍了转基因技术在汞污染土壤中的应用,探讨了转基因技术在环境治理和环境修复中潜在的应用价值.     

蚯蚓在植物修复芘污染土壤中的作用

采用盆栽试验法,研究了蚯蚓(Pheretima hupeiensis)在植物修复芘污染土壤中的作用。结果显示,试验浓度(20.24-321.42 mg/kg) 范围内,蚯蚓活动促进了芘污染土壤中修复植物黑麦草(Lolium multiforum)黑麦草的生长,其根冠比明显增大。添加蚯蚓72 d后,种植黑麦草的土壤中芘的去除率高达60.01%-86.26%,其平均去除率(74.66%)比无蚯蚓活动的土壤-植物系统(64.55%)提高10.11%,比无植物对照组(18.24%)提高56.42%。各种生物、非生物修复因子中,植物-微生物交互作用对芘去除的平均贡献率(51.75%)最为突出,比无蚯蚓活动时(44.94%)提高6.81%。说明蚯蚓活动可强化土壤-植物系统对土壤芘污染的修复作用。     

土壤重金属污染的植物修复

土壤重金属污染的危害范围广泛,使用传统的物理和化学修复方法成本高,对环境扰动大,而利用植物修复的效果较为明显,易于操作.本文论述了土壤重金属污染的单一植物、植物与微生物联合、植物与化学方法相结合.的修复方法,着重介绍了重金属超富'集植物的研究和植物体内螯合肽(PCs)的合成.生物螯合剂的应用及土壤重金属污染的动物、植物和微生物的联合修复将是未来研究的热点.     

Use of integrated phytoremediation for cleaning-up of oil-sludge-contaminated soil

The possibility of using multicomponent systems, including plants, mineral fertilizers, and plant growth promoting microorganisms, has been studied in vegetative experiments in order to stimulate phytoremediation of oil-sludge-contaminated soil. Winter rye (Secale cereale L.) was used as the principal phytoremediating plant species, whereas alfalfa (Medicago sativa L.), nitrogen fertilizer (ammonium nitrate), and a PGPR strain (Azospirillum brasilense SR80) were applied as additional components, individually or in various combinations. The obtained data revealed the critical importance of alfalfa for phytoremediation of hydrocarbon-contaminated soil. Application of different multicomponent treatments resulted in approximately 70% reduction of pollutant concentration in soil. The developed technological approaches were successfully tested in the remediation of an ex-oil-sludge pit on the ground of a petroleum refinery.     

The phytoremediation of an organic and inorganic polluted soil: A real scale experience

A phytoremediation process with horse manure, plants (Populus alba, Cytisus scoparius, Paulownia tomentosa) and naturally growing vegetation was carried out at a real-scale in order to phytoremediate and functionally recover a soil contaminated by metals (Zn, Pb, Cd, Ni, Cu, Cr), hydrocarbons (TPH) and polychlorobiphenyls (PCB).

All the plants were effective in two years in the reclamation of the polluted soil, showing an average reduction of about 35%, 40%, and 70% in metals, TPH and PCB content, respectively. As regards the plants, the poplar contributed the most to organic removal. In fact, its ability to take up and detoxify organic pollutants is well known. Paulownia tomentosa, instead, showed high metal removal. The Cytisus scoparius was the least effective plant in soil decontamination. The recovery of soil functionality was followed by enzyme activities, expressing the biochemical processes underway, and nutrient content useful for plant growth and development. Throughout the area, an enhancement of metabolic processes and soil chemical quality was observed. All the enzymatic activities showed a general increase over time (until 3-4 fold than the initial value for urease and β-glucosidase). Moreover, Cytisus scoparius, even though it showed a lower decontamination capability, was the most effective in soil metabolic stimulation.     

Influence of biochar and compost on phytoremediation of oil-contaminated soil

The use of pyrolyzed carbon, biochar, as a soil amendment is of potential interest for improving phytoremediation of soil that has been contaminated by petroleum hydrocarbons. To examine this question, the research reported here compared the effects of biochar, plants (mesquite tree seedlings), compost and combinations of these treatments on the rate of biodegradation of oil in a contaminated soil and the population size of oil-degrading bacteria. The presence of mesquite plants significantly enhanced oil degradation in all treatments except when biochar was used as the sole amendment without compost. The greatest extent of oil degradation was achieved in soil planted with mesquite and amended with compost (44% of the light hydrocarbon fraction). Most probable number assays showed that biochar generally reduced the population size of the oil-degrading community. The results of this study suggest that biochar addition to petroleum-contaminated soils does not improve the rate of bioremediation. In contrast, the use of plants and compost additions to soil are confirmed as important bioremediation technologies.     

PAHs污染土壤植物修复对酶活性的影响

PAHs作为一类持久性有机污染物对土壤环境质量产生深刻的影响。选用了中国亚热带城市普遍采用的4个树种(樟树、栾树、广玉兰、马褂木),利用盆栽试验,研究了PAHs污染土壤植物修复对酶活性影响。结果表明,多酚氧化酶活性定量抑制率为-94.98%—16.29%,过氧化氢酶为-76.71%—13.19%,磷酸酶为-49.62%—56.38%。土壤酶活性对PAHs污染的响应受到不同树种的影响。方差分析表明,过氧化氢酶活性在不同污染水平间差异显著,3种酶活性在不同时间下差异性显著,3种酶活性在不同树种×污染水平、不同时间×污染水平二因素作用下差异都不显著。主成分分析表明,PAHs污染对土壤酶活性的影响大于树种的影响,多酚氧化酶和磷酸酶对土壤反映敏感。     

Electrokinetic-enhanced phytoremediation of uranium-contaminated soil using sunflower and Indian mustard

Abstract

Electrokinetic-enhanced phytoremediation is an effective technology to decontaminate heavy metal contaminated soil. In this study, we examined the effects of electrokinetic treatments on plant uptake and bioaccumulation of U from soils with various U sources. Redistribution of uranium in soils as affected by planting and electrokinetic treatments was investigated. The soil was spiked with 100?mg kg^–1 UO₂, UO₃, and UO₂(NO₃)₂. After sunflower and Indian mustard grew for 60 days, 1 voltage of direct-current was applied across the soils for 9 days. The results indicated that U uptake in both plants were significantly enhanced by electrokinetic treatments from soil with UO₃ and UO₂(NO₃)₂. U was more accumulated in roots than in shoots. Electrokinetic treatments were effective on lowering soil pH near the anode region. Overall, uranium (U) removal efficiency reached 3.4–4.3% from soils with UO₃ and uranyl with both plants while that from soil with UO₂ was 0.7–0.8%. Electrokinetic remediation treatment significantly enhanced the U removal efficiency (5–6%) from soils with UO₃ and uranyl but it was 0.8–1.3% from soil with UO₂, indicating significant effects of U species and electrokinetic enhancement on U bioaccumulation. This study implies the potential feasibility of electrokinetic-enhanced phytoremediation of U soils with sunflower and Indian mustard.     

微生物对植物修复重金属污染土壤的促进效果

以印度芥菜作为超富集植物,通过盆栽试验研究了巨大芽胞杆菌和胶质芽胞杆菌的混合微生物制剂、黑曲霉30177发酵液对植物修复Cd、Pb、Zn污染土壤的作用.结果表明：巨大芽胞杆菌和胶质芽胞杆菌的混合微生物制剂不仅可以促进超富集植物的生长,增强超富集植物对土壤Cd、Pb、Zn的吸收,而且大幅度提高了植物的修复效率,在添加外源可溶性Cd、Pb、Zn的污染土壤上,可分别使印度芥菜提取量(以植物干质量计)提高1.18、1.54和0.85倍,在添加底泥Cd、Pb、Zn污染的土壤上,可分别使印度芥菜提取量提高4.00、0.64和0.65倍,在底泥污染的土壤上的促进效果明显强于外源添加污染的土壤.黑曲霉30177发酵液能显著促进印度芥菜对土壤Cd、Pb、Zn的吸收,在添加外源可溶性Cd、Pb、Zn的污染土壤上,印度芥菜地上部Cd、Pb、Zn的吸收量分别比对照提高了88.82%、129.04%和16.80%;在添加底泥Cd、Pb、Zn污染的土壤上,可分别比对照提高78.95%、113.63%和33.85%;但它可导致印度芥菜生物量的大幅度降低,起不到提高植物修复提取量的效果.经反相高效液相色谱初步分析发现,胶质芽胞杆菌、巨大芽胞杆菌发酵液中含有草酸、柠檬酸等有机酸,有机酸对重金属有一定的溶解作用,从而提高了重金属的生物有效性.     

蚯蚓-菌根在植物修复镉污染土壤中的作用

以灰化土(Aquods)为供试土壤,分别加入4个浓度的Cd2 (0,5,10,20mg/kg)模拟土壤污染,设置每钵接种8条蚯蚓(Pheretimasp.)、接种菌根(InoculumEndorize-Mix2)和同时接种蚯蚓和菌根的处理,以不加蚯蚓和菌根为对照,并种植黑麦草(Loliummultiflorum),研究蚯蚓菌根相互作用对Cd污染土壤中黑麦草生长及土壤中Cd生物有效性的影响。结果表明菌根浸染率不受添加Cd浓度的影响,平均浸染率为22%,加入蚯蚓能使菌根的侵染率提高9%。在Cd污染土壤上,引进蚯蚓显著增加了黑麦草地上部的生物量,接种菌根对黑麦草地上部分产量没有明显影响,同时接种蚯蚓和菌根与只接种蚯蚓相比没有显著差异。蚯蚓活动显著提高了土壤中CaCl2-Cd的含量,而菌根只在低浓度Cd处理上增加了土壤中CaCl2-Cd含量,二者对H2O-Cd、DTPA-Cd均无显著影响,蚯蚓和菌根对增加土壤有效态Cd含量不存在协同作用。蚯蚓活动促进了黑麦草对Cd的吸收,但吸收的Cd积累于黑麦草根部。接种菌根不仅能促进黑麦草对Cd的吸收,而且还能促进Cd从植物的根部向地上部分转移,由于接种蚯蚓可以提高菌根的浸染率,所以二者具有促进Cd向地上部转移的协同作用。这对于重金属污染土壤的植物修复具有十分重要的意义。     

Optimizing phytoremediation of heavy metal-contaminated soil by exploiting plants' stress adaptation

Soil phytoextraction is based on the ability of plants to extract contaminants from the soil. For less bioavailable metals, such as Pb, a chelator is added to the soil to mobilize the metal. The effect can be significant and in certain species, heavy metal accumulation can rapidly increase 10-fold. Accumulation of high levels of toxic metals may result in irreversible damage to the plant. Monitoring and controlling the phytotoxicity caused by EDTA-induced metal accumulation is crucial to optimize the remedial process, i.e. to achieve maximum uptake. We describe an EDTA-application procedure that minimizes phytotoxicity by increasing plant tolerance and allows phytoextraction of elevated levels of Pb and Cd. Brassica juncea is tested in soil with typical Pb and Cd concentrations of 500 mg kg-1 and 15 mg kg-1, respectively. Instead of a single dose treatment, the chelator is applied in multiple doses, that is, in several small increments, thus providing time for plants to initiate their adaptation mechanisms and raise their damage threshold. In situ monitoring of plant stress conditions by chlorophyll fluorescence recording allows for the identification of the saturating heavy metal accumulation process and of simultaneous plant deterioration.     

Biotechnological applications of serpentine soil bacteria for phytoremediation of trace metals

Serpentine or ultramafic soils are produced by weathering and pedogenesis of ultramafic rocks that are characterized by high levels of Ni, Cr, and sometimes Co, but contain low levels of essential nutrients such as N, P, K, and Ca. A number of plant species endemic to serpentine soils are capable of accumulating exceptionally high concentrations of Ni, Zn, and Co. These plants are known as metal “hyperaccumulators.” The function of hyperaccumulation depends not only on the plant, but also on the interaction of the plant roots with rhizosphere microbes and the concentrations of bioavailable metals in the soil. The rhizosphere provides a complex and dynamic microenvironment where microorganisms, in association with roots, form unique communities that have considerable potential for the detoxification of hazardous materials. The rhizosphere bacteria play a significant role on plant growth in serpentine soils by various mechanisms, namely, fixation of atmospheric nitrogen, utilization of 1-aminocyclopropane-1-carboxylic acid (ACC) as the sole N source, production of siderophores, or production of plant growth regulators (hormones). Further, many microorganisms in serpentine soil are able to solubilize “unavailable” forms of heavy metal–bearing minerals by excreting organic acids. In addition, the metal-resistant serpentine isolates increase the efficiency of phytoextraction directly by enhancing the metal accumulation in plant tissues and indirectly by promoting the shoot and root biomass of hyperaccumulators. Hence, isolation of the indigenous and stress-adapted beneficial bacteria serve as a potential biotechnological tool for inoculation of plants for the successful restoration of metal-contaminated ecosystems. In this study, we highlight the diversity and beneficial features of serpentine bacteria and discuss their potential in phytoremediation of serpentine and anthropogenically metal-contaminated soils.     

一种新发现的铜积累植物——密毛蕨

对铜尾矿上生长的密毛蕨(Petridium revolutum)进行了野外调查分析和温室营养液砂培实验.结果表明,密毛蕨所生长的废铜矿土壤中Cu含量平均为2 432 mg·kg^-1 DW,最高达7 554 mg·kg^-1 DW;地上部生物量平均为18.33 g·plant^-1 DW,最高达40.05 g·plant^-1 DW;地上部Cu含量平均为201 mg·kg^-1 DW,最高达567 mg·kg^-1 DW;地下部Cu含量平均为346 mg·kg^-1 DW,最高达1 723 mg kg^-1 DW;密毛蕨对Cu的转移系数平均为0.81,最高达3.88.在营养液砂培的条件下,Cu 7 mg·L^-1处理没有抑制密毛蕨地上部的生长;密毛蕨体内的Cu含量随着介质中Cu浓度的增加而显著增加,但是大部分的Cu积累在地下部.密毛蕨对Cu具有较强的忍耐和较高的积累能力,可作为修复Cu污染土壤的新材料.